Chromophoric polymer dots with narrow-band emission

ABSTRACT

Polymers, monomers, chromophoric polymer dots and related methods are provided. Highly fluorescent chromophoric polymer dots with narrow-band emissions are provided. Methods for synthesizing the chromophoric polymers, preparation methods for forming the chromophoric polymer dots, and biological applications using the unique properties of narrow-band emissions are also provided.

CROSS REFERENCES TO RELATED APPLICATIONS

The present application is a U.S. National Phase Application under 35U.S.C. § 371 of International Application No. PCT/US2012/071767, filedDec. 27, 2012, which claims priority to U.S. Provisional Application No.61/582,181, filed Dec. 30, 2011, and U.S. Provisional Application No.61/607,455, filed on Mar. 6, 2012, the entirety of each of which isincorporated by reference herein.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

This invention was made with government support under grant numberCA147831 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND OF THE INVENTION

Recent developments in fluorescence detection coupled withbioconjugation techniques are leading to a rapid proliferation ofadvanced fluorescence-based techniques in chemistry and the lifesciences, such as fluorescence microscopy, flow cytometry, versatilebiological assays, and biosensors. These fluorescence techniques makeextensive use of organic dye molecules as probes. However, intrinsiclimitations of the conventional dyes, such as low absorptivity and poorphotostability, have posed difficulties in further developments ofhigh-sensitivity imaging techniques and high-throughput assays. A numberof strategies for developing brighter fluorescent probes have beenpursued. For example, luminescent nanoparticles such as inorganicsemiconductor quantum dots (Qdots) are under active development and nowcommercially available from Life Technologies (Invitrogen). (Bruchez,M.; Moronne, M.; Gin, P.; Weiss, S.; Alivisatos, A. P, Science 1998,281, 2013-2016. Michalet, X.; Pinaud, F. F.; Bentolila, L. A.; Tsay, J.M.; Doose, S.; Li, J. J.; Sundaresan, G.; Wu, A. M.; Gambhir, S. S.;Weiss, S. Science 2005, 307, 538-544.) An alternative fluorescentnanoparticle is dye doped latex spheres, which exhibit improvedbrightness and photostability as compared to single fluorescentmolecules because of multiple dye molecules per particle and theprotective latex matrix. (Wang, L.; Wang, K. M.; Santra, S.; Zhao, X.J.; Hilliard, L. R.; Smith, J. E.; Wu, J. R.; Tan, W. H. Anal. Chem.2006, 78, 646-654).

The limitations of current luminescent particles provide a need forexploring alternative strategies for the design of more highlyfluorescent nanoparticles. Recently, fluorescence semiconducting polymerdots (Pdots) have attracted interest because of their fluorescencebrightness and photostability as compared to Qdots and dye-loaded latexbeads. The use of fluorescent polymer dots as fluorescent probes alsocan confer other useful aspects. Recently, surface functionalization hasbeen achieved by a co-condensation scheme where amphiphilic polymermolecules bearing functional groups were blended with semiconductingpolymers to form Pdots with surface reactive groups. Bioconjugation hasbeen demonstrated by reacting the functional groups with biomolecules,and the Pdot-bioconjugates can specifically and effectively labelbiomolecules for cellular imaging, bioorthogonal labeling, and in vivotumor targeting.

However, there can be drawbacks when the current Pdots are used asfluorescent probes in practical applications. Many biologicalapplications can include detecting multiple targets simultaneously;thus, there is a need for probes that possess narrow-band emission peaksfor spectral multiplexing. However, currently available Pdots canexhibit very broad emission spectra, which limit their usefulness inpractical applications. The spectral width of a fluorescent probe can becharacterized by the full width at half maximum (FWHM) of its emissionpeak. In general, currently available Pdots exhibit broad emissionspectra with large FWHMs. Such broad emission spectra are a drawback formulti-target detection in biology. Therefore, there is a need to designand develop new type of Pdots with narrow-band emissions.

BRIEF SUMMARY OF THE INVENTION

The present invention provides chromophoric polymers that can form, forexample, highly fluorescent chromophoric polymer dots with narrow-bandemissions. The present invention also provides design considerations insynthesizing these chromophoric polymers, preparation methods forforming the relative polymer dots, and biological applications using theunique properties of narrow-band emissions. Chromophoric polymer dotswith narrow-band emissions bring forward unique properties of highlyfluorescent nanoparticle bioconjugates for a wide range offluorescence-based applications.

In one aspect, the present invention provides chromophoric polymer dotswith narrow-band emissions. The emission wavelength of the polymer dotscan vary from ultraviolet to near infrared region. The full width athalf maximum (FWHM) of the emission band is less than 70 nm. In someembodiments, the FWHM is less than about 65 nm. In some embodiments, theFWHM is less than about 60 nm. In some embodiments, the FWHM is lessthan about 55 nm. In some embodiments, the FWHM is less than about 50nm. In some embodiments, the FWHM is less than about 45 nm. In someembodiments, the FWHM is less than about 40 nm. In some embodiments, theFWHM is less than about 35 nm. In some embodiments, the FWHM is lessthan about 30 nm. In some embodiments, the FWHM is less than about 25nm. In some embodiments, the FWHM is less than about 20 nm. In someembodiments, the FWHM is less than about 10 nm. In some embodiments, theFWHM of the polymer dots described herein can range between about 5 nmto about 70 nm, from about 10 nm to about 60 nm, from about 20 nm toabout 50 nm, or from about 30 nm to about 45 nm.

In some embodiments, the narrow-band emissive Pdots include at least onechromophoric polymer. The narrow-band emissive Pdots can also include anarrow-band emissive unit covalently attached to the chromophroicpolymer, where the narrow-band emissive unit gives narrow-bandemissions. The narrow-band emissive unit can be incorporated into thepolymer backbone. The narrow-band emissive unit can also be covalentlyattached to the side chain, or terminal unit of the polymer. Thenarrow-band emissive Pdots can also include chromophoric polymer dotsdoped with inorganic materials, where the inorganic materials givenarrow-band emissions. The narrow-band emissive Pdots can include onlychromophoric polymers, where the chromophoric polymers give narrow-bandemissions. The emission FWHM of the above Pdots is less than 70 nm. Incertain embodiments, the FWHM is less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive Pdots include chromophoricpolymer dots chemically cross-linked with other narrow-band species suchas dyes (e.g., polymer or small molecule dyes). The narrow-band emissivePdots can include only chromophoric polymers that give narrow-bandemissions. The emission FWHM of the above Pdots is less than 70 nm. Incertain embodiments, the FWHM is less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymer dots include atleast one narrow-band emissive chromophoric polymers. The narrow-bandemissive polymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotscan include narrow-band monomers. The narrow-band emissive polymer dotscan also include any other monomers. The narrow-band monomers can beenergy acceptors so that the final Pdots can exhibit narrow-bandemissions. The narrow-band emissive chromophoric polymers in goodsolvents may exhibit broad emissions or narrow emissions. However, theirnanoparticle form gives narrow-band emissions. The emission FWHM of theabove Pdots is less than 70 nm. In certain embodiments, the FWHM is lessthan 60 nm, less than 50 nm, less than 40 nm, less than 30 nm, or lessthan 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude boron-dipyrromethene(4,4-difluoro-4-bora-3a,4a-diaza-s-indacene, BODIPY) based monomers andtheir derivatives as narrow-band monomers. BODIPY monomers and theirderivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, BODIPY extended systems and BODIPYanalogues. The narrow-band emissive polymers can also include any othermonomers. The BODIPY based-monomers can be energy acceptors so that thefinal Pdots can exhibit narrow-band emissions. The narrow-band emissivechromophoric polymers in good solvents may exhibit broad emissions ornarrow emissions. However, their nanoparticle form gives narrow-bandemissions. The emission FWHM of the above Pdots is less than 70 nm. Incertain embodiments, the FWHM is less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude squaraine and their derivatives as narrow-band monomers.Squaraine derivatives include but are not limited to their alkylderivatives, aryl derivatives, alkyne derivatives, aromatic derivatives,alkoxide derivatives, aza derivatives, their extended systems andanalogues. The narrow-band emissive polymers can also include any othermonomers. The squaraine and their derivatives can be energy acceptors sothat the final Pdots can exhibit narrow-band emissions. The narrow-bandemissive chromophoric polymers in good solvents may exhibit broademissions or narrow emissions. However, their nanoparticle form givesnarrow-band emissions. The emission FWHM of the above Pdots is less than70 nm. In certain embodiments, the FWHM is less than 60 nm, less than 50nm, less than 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude metal complexes and their derivatives as narrow-band monomers.Metal complexes and their derivatives include but are not limited totheir alkyl derivatives, aryl derivatives, alkyne derivatives, aromaticderivatives, alkoxide derivatives, aza derivatives, their extendedsystems and analogues. The narrow-band emissive polymers can alsoinclude any other monomers. The metals can be any metal such as Na, Li,Zn, Mg, Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt, Pd, Ru, Rh, Re, Os, Ir,Ag, Au and so on. The metal complexes can be energy acceptors so thatthe final Pdots can exhibit narrow-band emissions. The narrow-bandemissive chromophoric polymers in good solvents may exhibit broademissions or narrow emissions. However, their nanoparticle form givesnarrow-band emissions. The emission FWHM of the above Pdots is less than70 nm. In certain embodiments, the FWHM is less than 60 nm, less than 50nm, less than 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude porphyrin, metalloporphyrin, and their derivatives asnarrow-band monomers. Porphyrin, metalloporphyrin, and their derivativesinclude but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Themetals in the metalloporphyrins can be any metal such as Na, Li, Zn, Mg,Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt, Pd, Ru, Rh, Re, Os, Ir, Ag, Auand so on. The narrow-band emissive polymers can also include any othermonomers. The porphyrin, metalloporphyrin and their derivatives can beenergy acceptors so that the final Pdots can exhibit narrow-bandemissions. The narrow-band emissive chromophoric polymers in goodsolvents may exhibit broad emissions or narrow emissions. However, theirnanoparticle form gives narrow-band emissions. The emission FWHM of theabove Pdots is less than 70 nm. In certain embodiments, the FWHM is lessthan 60 nm, less than 50 nm, less than 40 nm, less than 30 nm, or lessthan 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude phthalocyanine and its derivatives as narrow-band monomers.Phthalocyanine derivatives include but are not limited to their alkylderivatives, aryl derivatives, alkyne derivatives, aromatic derivatives,alkoxide derivatives, aza derivatives, their extended systems andanalogues. The metals in the phthalocyanine derivatives can be any metalsuch as Na, Li, Zn, Mg, Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt, Pd, Ru,Rh, Re, Os, Ir, Ag, Au and so on. The narrow-band emissive polymers canalso include any other monomers. The phthalocyanine derivatives can beenergy acceptors so that the final Pdots can exhibit narrow-bandemissions. The narrow-band emissive chromophoric polymers in goodsolvents may exhibit broad emissions or narrow emissions. However, theirnanoparticle form gives narrow-band emissions. The emission FWHM of theabove Pdots is less than 70 nm. In certain embodiments, the FWHM is lessthan 60 nm, less than 50 nm, less than 40 nm, less than 30 nm, or lessthan 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude Lanthanide complexes and their derivatives as narrow-bandmonomers. Lanthanide complexes and their derivatives include but are notlimited to their alkyl derivatives, aryl derivatives, alkynederivatives, aromatic derivatives, alkoxide derivatives, azaderivatives, their extended systems and analogues. The narrow-bandemissive polymers can also include any other monomers. The Lanthanidecomplexes and their derivatives can be energy acceptors so that thefinal Pdots can exhibit narrow-band emissions. The narrow-band emissivechromophoric polymers in good solvents may exhibit broad emissions ornarrow emissions. However, their nanoparticle form gives narrow-bandemissions. The emission FWHM of the above Pdots is less than 70 nm. Incertain embodiments, the FWHM is less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude perylene and its derivatives as narrow-band monomers. Perylenederivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Theperylene derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude cyanine and its derivatives as narrow-band monomers. Cyaninederivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Thecyanine derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude rhodamine and its derivatives as narrow-band monomers. Rhodaminederivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Therhodamine derivatives can be energy acceptors so that the final Pdotscan exhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude coumarin and its derivatives as narrow-band monomers. Coumarinderivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Thecoumarin derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude xanthene and its derivatives as narrow-band monomers. Xanthenederivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Thexanthene derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

In another aspect, the present invention provides functionalizedchromophoric polymer dot. The functionalized Pdot include a narrow-bandemissive Pdot and a functional group that is physically or chemicallyattached to the Pdot.

In yet another aspect, the present invention discloses a bioconjugate ofthe polymer dots with narrow-band emissions. The bioconjugate is formedby the attachment of a biomolecule to one or more functional groups ofthe narrow-band emissive chromophoric polymer dot. The attachment may bedirect or indirect.

In yet another aspect, methods of preparing narrow-band emissivechromophoric polymer dots are disclosed. In some embodiments, thesechromophoric polymer dots can be formed using nanoprecipitation. Thenanoprecipitation method involves the introduction of a solution of apolymer in a good solvent into a poor solvent, where the solubilitycollapse the polymer into a nanoparticle form. In certain embodiments,the chromophoric polymer with narrow-band emissions can be preparedusing the mini-emulsion method.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows example schematic structures of narrow-band emissivepolymers. FIG. 1A shows the structure of a homopolymer that comprisesonly one narrow-band monomer. FIG. 1B shows the structure of a two-unitcopolymer that includes one narrow-band monomer and one general monomer.The narrow-band monomer can be an energy-acceptor and the generalmonomer can be an energy-donor. Energy-transfer inside Pdots can resultin narrow-band emissions. FIG. 1C shows the structure of a three-unitcopolymer that includes one narrow-band monomer and two general monomerssuch as general monomer 1 (D1) and general monomer 2 (D2). Thenarrow-band monomer can be an energy-acceptor, general monomer 1 can bean energy-donor, general monomer 2 can also be a donor to thenarrow-band monomer. In some embodiments, general monomer 2 can besimultaneously an energy-acceptor from monomer 1 and an energy-donor tothe narrow-band monomer. Multi-step energy-transfer inside Pdots canresult in narrow-band emissions. FIG. 1D shows the structure of atwo-unit copolymer that includes the narrow-band unit cross-linked withthe side-chains. The copolymer backbone can be an energy-donor, and thenarrow-band unit can be an energy-acceptor. Energy-transfer inside Pdotscan result in narrow-band emissions. FIG. 1E shows the structure of ahomopolymer that includes the narrow-band unit cross-linked with theside-chains. The homopolymer backbone can be an energy-donor, and thenarrow-band unit can be an energy-acceptor. Energy-transfer inside Pdotscan result in narrow-band emissions. FIG. 1F shows a structure of apolymer that includes a narrow-band unit attached to a terminus of thepolymer. The polymer backbone can be an energy-donor, and thenarrow-band unit can be an energy-acceptor. Energy-transfer inside Pdotscan result in narrow-band emissions. FIG. 1G-1L show example schematicstructures of narrow-band emissive polymers that include a generalmonomer, a narrow-band monomer, and a functional monomer (or afunctional group). The functional monomer can, e.g., provide reactivechemical group for chemical reactions or bioconjugation reactions.

FIGS. 2A-2N show non-limiting examples of the chemical structures ofnarrow-band emissive copolymers. The copolymer can include one generalmonomer as an energy donor and one narrow-band monomer as an acceptor.FIG. 2A shows a non-limiting list of examples of the general monomers.FIGS. 2B-2N shows a non-limiting list of examples of the narrow-bandemissive copolymer including one general monomer chosen from FIG. 2A anddifferent BODIPY derivatives or other boron-containing units asnarrow-band monomers. FIGS. 2M and 2N show some specific examples of thenarrow-band emissive copolymers based on the chemical structures inFIGS. 2B-2L.

FIGS. 3A-E show a non-limiting list of examples of the chemicalstructures of general D1 type monomers and D2 type monomers used forsynthesizing narrow-band emissive polymers, e.g., as in FIG. 1 and FIG.31A. FIG. 3A shows example D1 monomers. FIGS. 3B, 3C, 3D and 3E showexample D2 monomers and example derivatives of D2 monomers. Thederivatives of D2 monomers are marked as D2′ monomers in the figures.The general D1 type monomers can, e.g., be copolymerized with the D2type (or D2′ type) and the narrow-band monomer to obtain narrow-bandemissive polymer. Any, e.g., of the D1 type monomers, D2 type, or D2′type monomers can also be separately used to copolymerize with onenarrow-band monomer to obtain the narrow-band emissive polymers as inFIG. 1. Rather than copolymerization, a narrow band emissive unit can,e.g., be attached to the side chains or termini of a polymer formed fromany of the D1 type monomers, D2 type, or D2′ type monomers.

FIGS. 4A-4H show a non-limiting list of different BODIPY derivatives assome specific examples as a narrow-band monomer. Each of the BODIPYderivatives can be used to synthesize a narrow-band emissivehomopolymer. Each of the BODIPY derivatives can also be copolymerizedwith any of the general polymers to synthesize a narrow-band emissivecopolymer. Each of the BODIPY derivatives can be used as a narrow-bandemissive unit to cross-link with the side-chains of conventionalsemiconducting polymers to form narrow-band emissive polymers.

FIG. 5 shows a non-limiting list of dipyrrin-metal derivatives and somespecific examples as narrow-band monomer. Each of the dipyrrin-metalderivatives can be used to synthesize a narrow-band emissivehomopolymer. Each of the dipyrrin-metal derivatives can also becopolymerized with any of the general polymers to synthesize anarrow-band emissive copolymer. Each of the dipyrrin-metal derivativescan be used as a narrow-band emissive unit to cross-link with theside-chains of conventional semiconducting polymers to form narrow-bandemissive polymers.

FIGS. 6A-6E show a non-limiting list of examples of squarainederivatives as narrow-band monomer. Each of the squaraine derivativescan be used to synthesize a narrow-band emissive homopolymer. Each ofthe squaraine derivatives can also be copolymerized with any of thegeneral polymers to synthesize a narrow-band emissive copolymer. Each ofthe squaraine derivatives can be used as a narrow-band emissive unit tocross-link with the side-chains of conventional semiconducting polymersto form narrow-band emissive polymers.

FIGS. 7A-7C show a non-limiting list of narrow-band emissive polymerincluding metal complexes and their derivatives as narrow-band monomer.Different Pt complexes were used as narrow-band monomers in the listedpolymers, and other metal complexes can also be used. Each of the metalcomplexes can be used to synthesize a narrow-band emissive homopolymer.Each of the metal complexes can also be copolymerized with any of thegeneral polymers to synthesize a narrow-band emissive copolymer. Each ofthe metal complexes can be used as a narrow-band emissive unit tocross-link with the side-chains of conventional semiconducting polymersto form narrow-band emissive polymers.

FIG. 8 shows a non-limiting list of narrow-band emissive polymerincluding porphyrin, metalloporphyrin and their derivatives asnarrow-band monomer. Each of the porphyrin derivatives can be used tosynthesize a narrow-band emissive homopolymer. Each of the porphyrinderivatives can also be copolymerized with any of the general polymersto synthesize a narrow-band emissive copolymer. Each of the porphyrinderivatives can be used as a narrow-band emissive unit to cross-linkwith the side-chains of conventional semiconducting polymers to formnarrow-band emissive polymers.

FIG. 9 shows a non-limiting list of phthalocyanine and its derivativesas narrow-band monomers. Each of the phthalocyanine derivatives can beused to synthesize a narrow-band emissive homopolymer. Each of thephthalocyanine derivatives can also be copolymerized with any of thegeneral polymers to synthesize a narrow-band emissive copolymer. Each ofthe phthalocyanine derivatives can be used as a narrow-band emissiveunit to cross-link with the side-chains of conventional semiconductingpolymers to form narrow-band emissive polymers.

FIG. 10A-10H shows a non-limiting list of narrow-band emissive polymercomprising lanthanide complexes and their derivatives as e.g.,narrow-band monomer. FIG. 10A shows example schematic structures ofnarrow-band emissive polymers that include lanthanide complexes asnarrow-band emissive units. FIG. 10B shows example structures ofeuropium (Eu) and terbium (Tb) complexes as narrow-band monomers in thelisted polymers. The D type monomers can be energy donors that transferenergy to lanthanide complex. FIG. 10C shows example chemical structuresof general polymers as donors that can transfer energy to lanthanidecomplexes in narrow-band emissive Pdots. FIG. 10D shows example chemicalstructures of anionic ligand (L1) to form lanthanide complexes. FIG. 10Eshows example chemical structures of the substituted groups in theligands (L1 and L2) of lanthanide complexes. FIG. 10F shows examplechemical structures of neutral ligands (L2) to form lanthanidecomplexes. FIG. 10G shows a schematic illustration of formingnarrow-band emissive polymers that include general polymer as donors andthe lanthanide complex as narrow-band emissive units. Both the generalpolymer and lanthanide complexes include amino groups which can be,e.g., covalently cross-linked with an amine-reactive polymer to form thelanthanide-complex grafted polymers for preparing narrow-band emissivePdots. FIG. 10H shows schematic illustration of forming narrow-bandemissive polymers that include only the lanthanide complex asnarrow-band emissive units. The lanthanide complexes include aminogroups which can be, e.g., covalently cross-linked with anamine-reactive polymer to form the lanthanide-complex grafted polymersfor preparing narrow-band emissive Pdots.

FIG. 11A shows a non-limiting list of perylene and its derivatives asnarrow-band monomer. Each of the perylene derivatives can be used tosynthesize a narrow-band emissive homopolymer. Each of the perylenederivatives can also be copolymerized with any of the general polymersto synthesize a narrow-band emissive copolymer. Each of the perylenederivatives can be used as a narrow-band emissive unit to cross-linkwith the side-chains of conventional semiconducting polymers to formnarrow-band emissive polymers. FIGS. 11B-11D show a non-limiting list ofcyanine and its derivatives as narrow-band monomer. Each of the cyaninederivatives can be used to synthesize a narrow-band emissivehomopolymer. Each of the cyanine derivatives can also be copolymerizedwith any of the general polymers to synthesize a narrow-band emissivecopolymer. Each of the cyanine derivatives can be used as a narrow-bandemissive unit to cross-link with the side-chains of conventionalsemiconducting polymers to form narrow-band emissive polymers.

FIG. 12A shows chemical structures of examples of BODIPY monomerssynthesized. FIG. 12B shows their absorption spectra, fluorescencespectra, and fluorescence quantum yield.

FIG. 13A shows a series of copolymers that include a general monomerfluorene and a narrow-band monomer (BODIPY Monomer 2a in FIG. 12) atdifferent molar ratios. FIG. 13B shows the fluorescence spectra of thepolymers in a good solvent tetrahydrofuran (THF). FIG. 13C shows thefluorescence spectra of the Pdots in water. As can be seen, the emissionspectra of the polymers in THF exhibit similar FWHM for all thepolymers. However, the Pdots shows quite different FWHM because of thechromophore packing in the nanoparticles. Narrow-band emissions can beobtained by adjusting the BODIPY ratio relative to the general fluorenemonomer.

FIG. 14A shows the chemical structure of a narrow-band emissive polymer(Polymer510) synthesized by using the BODIPY monomer 1a in FIG. 12 asnarrow-band monomer and several general monomers. FIG. 14B shows theabsorption spectrum and emission spectra of the Polymer510 Pdots inwater. When excited at 380 nm, the Pdot emission exhibits a FWHM of 41nm. When excited at 470 nm, the Pdot emission exhibits a FWHM of 25 nm.The fluorescence quantum yield was measured to be 64%.

FIG. 15A shows the chemical structure of a narrow-band emissive polymer(Polymer 590) synthesized by using the BODIPY monomer 2a in FIG. 12 asnarrow-band monomer and several general monomers. FIG. 15B shows theabsorption and emission spectra of the Polymer590 in THF. FIG. 15C showsthe absorption and emission spectra of the Polymer590 Pdots in water.The Pdot emission exhibits a FWHM of 64 nm and fluorescence quantumyield is 0.13. As indicated by the spectra, the polymer shows broademission in a good solvent such as THF. However, the Pdots shownarrow-band emission.

FIG. 16A shows the chemical structure of a narrow-band emissive polymer(Polymer 680) synthesized by using the BODIPY monomer 3a in FIG. 12 asnarrow-band monomer and several general monomers. FIG. 16B shows theabsorption and emission spectra of the Polymer680 in THF. FIG. 16C showsthe absorption and emission spectra of the Polymer680 Pdots in water.The Pdot emission exhibits a FWHM of 55 nm and fluorescence quantumyield is 0.19. As indicated by the spectra, the polymer shows broademission in a good solvent such as THF. However, the Pdots shownarrow-band emission.

FIG. 17 shows particle size distributions of narrow-band emissivePolymer510 Pdots, Polymer590 Pdots, and Polymer680 Pdots, respectively.The data were measured by dynamic light scattering.

FIGS. 18A-C show flow cytometry results of Polymer590 Pdots andPolymer680 Pdots labeled MCF-7 cells, respectively. FIG. 18A shows theside scattering (SSC) versus forward scattering (FSC). FIG. 18B showsthe fluorescence intensity distributions of the MCF-7 cells labeled withPolymer590 Pdots. Blue curve is the negative control, and orange curveis the positive labeling. FIG. 18C shows the fluorescence intensitydistributions of the MCF-7 cells labeled with Polymer680 Pdots. Greencurve is the negative control, and red curve is the positive labeling.

FIG. 19A shows fluorescence images of MCF-7 breast-cancer cells labeledwith Polymer590 Pdot-streptavidin. Negative labeling performed under thesame condition but in the absence of the biotinylated primary antibodydoes not show fluorescence signal. Images of blue fluorescence from thenuclear stain Hoechst 34580; orange fluorescence images from Pdot;Nomarski (DIC) images; combined fluorescence images. FIG. 19B showsfluorescence images of MCF-7 breast-cancer cells labeled with Polymer680Pdot-streptavidin. Negative labeling performed under the same conditionbut in the absence of the biotinylated primary antibody does not showfluorescence signal. Images of blue fluorescence from the nuclear stainHoechst 34580; red fluorescence images from Pdot; Nomarski (DIC) images;combined fluorescence images.

FIG. 20 shows multi-step synthesis of a series of copolymers thatinclude a general monomer fluorene and a narrow-band monomer (BODIPYMonomer 2a in FIG. 12) at different molar ratios.

FIG. 21 shows multi-step synthesis of the BODIPY monomer 1a in FIG. 12and the narrow-band emissive polymer Polymer510.

FIG. 22 shows multi-step synthesis of the BODIPY monomer 2a in FIG. 12and the narrow-band emissive polymer Polymer590.

FIG. 23 shows multi-step synthesis of the BODIPY monomer 3a in FIG. 12and the narrow-band emissive polymer Polymer680.

FIG. 24A shows multi-step synthesis of a squaraine derivative 1 and anarrow-band emissive polymer PFS (Polymer690) using a squarainederivative as a narrow-band monomer and fluorene as general monomer.FIG. 24B shows multi-step synthesis of a squaraine derivative 5 and thenarrow-band emissive polymer PFS5.5 using the squaraine as narrow-bandmonomer and fluorene as general monomer. FIG. 24C shows a schematicillustration of the squaraine based narrow emissive polymer dots andPdot-bioconjugates for specific cellular targeting.

FIG. 25A shows photophysical data of the narrow-band emissivefluorene-4% squaraine copolymer and fluorene-19% squaraine copolymer.FIG. 25B shows absorption spectra data of the fluorene-4% squarainecopolymer and fluorene-19% squaraine copolymer. FIG. 25C shows emissionspectra of the fluorene-4% squaraine copolymer and fluorene-19%squaraine copolymer excited at 405 nm. The fluorene-4% squarainecopolymer Pdots shows emission FWHM of 37 nm at 690 nm and fluorescencequantum yield is 0.23. FIG. 25D shows emission spectra of thefluorene-4% squaraine copolymer and fluorene-19% squaraine copolymerexcited at 675 nm. FIG. 25E shows photophysical data and particle sizeof the narrow-band emissive PFS Pdots and PFS 5.5 Pdots with varyingsquaraine ratio. FIG. 25F shows the fluorescence emission spectra of thenarrow-band emissive PFS Pdots and PFS 5.5 Pdots with varying squaraineratio. The PFS-1.5% squaraine copolymer Pdots shows emission FWHM of 37nm at 690 nm and fluorescence quantum yield is 0.30. FIG. 25G showsabsorption and fluorescence spectra of PFS and PFS5.5 Pdots with 1.5%squaraine molar ratio. Top-left panel a shows the absorption andfluorescence spectra of PFS Pdot with 1.5% squaraine molar ratio.Top-right panel b shows the fluorescence spectra of PFS Pdot at 1.5%molar ratio of squaraine dye in THF and the Pdots formed in water.Bottom-left panel c shows the absorption spectra of PFS and PFS 5.5Pdots (with 1.5% squaraine molar ratio) in water. Bottom-right panel dshows the fluorescence spectra of PFS and PFS 5.5 Pdots (with 1.5%squaraine molar ratio) in water. FIG. 25H shows the particle sizedistributions of PFS Pdots (average size of 19 nm) and PFS 5.5 Pdots(average size of 19 nm). Bottom panels show the TEM images of PFS Pdots(c) and PFS 5.5 Pdots (d). FIG. 25I shows single-particle brightnessimages with 405 nm excitation for three samples: (a) Qdots 705; (b) 1.5%PFS Pdots; (c) 1.5% PFS 5.5 Pdots. The images in the top panel wereobtained under identical excitation and detection conditions. All scalebars represent 5 μm. The bottom panels show the histograms of thebrightness distributions. FIG. 25J shows flow cytometry intensitydistributions of MCF-7 breast cancer cells labeled with Qdot705-streptavidin, PFS Pdot-streptavidin, and PFS5.5 Pdot-streptavidin.FIG. 25K shows confocal fluorescence images of MCF-7 cells labeled withPFS Pdot-streptavidin and PFS 5.5 Pdot-streptavidin probes.

FIG. 26A shows the scheme for the preparation of Pdot-Qdot hybrid NPs.PFBT with amino terminal groups was converted to thiols first in orderto covalently bind to the surfaces of QD. Then Pdot-Qdot mixtures weremixed well with PS-PEG-COOH in THF following the nanoprecipitation inwater under vigorous sonication to make QD-embedded Pdots. FIG. 26Bshows TEM images of Pdot-Qdot nanocomposites. The inset in theupper-left corner shows the enlarged view of single Pdot-Qdotnanocomposites. The blue and white scale bars represent 20 nm and 2 nm,respectively. FIG. 26C shows DLS measurements of the hydrodynamicdiameters of Pdot-Qdot NPs.

FIG. 27A shows UV-visible spectrum of Pdot-Qdot NPs in water. FIG. 27Bshows emission spectra of the Pdot-Qdot nanocomposites (solid lines).Dashed spectra show emission from QD655 (red), QD705 (purple), and QD800(pink) in decane. FIG. 27C shows single-particle fluorescence images ofPFBT-DBT Pdots (upper graph) and the corresponding intensitydistribution histograms (bottom graph). FIG. 27D shows single-particlefluorescence images of PFBT-QD655 Pdots (upper graph) and thecorresponding intensity distribution histograms (bottom graph). Thescale bars are 4 μm.

FIGS. 28A-28C show the two-color confocal microscopy images ofmicrotubules in HeLa cells labeled with Pdot-QD705-streptavidin. Theblue fluorescence is from nuclear counter-stain Hoechst 34580 (A), thered fluorescence (B) is from Pdot-QD705-streptavidin, and (C) is theoverlay of panels (A) and (B). FIGS. 28D-28F are images of the controlsample where cells were incubated with Pdot-QD705-streptavidin but inthe absence of biotinylated primary antibody. The scale bars are 20 μm.FIGS. 28G-28H show flow cytometry results of Pdot-Qdot labeled MCF-7cells. The purple and pink lines show the fluorescence intensitydistributions of Pdot-QD705-streptavidin and Pdot-QD800-streptavidinlabeled cells, respectively. The black lines represent the results ofcontrol samples (no primary Biotin anti-human CD326 EpCAM antibody).

FIG. 29A shows the chemical structure of a conventional broad-bandemission semiconducting polymer PFBT and a narrow-band emissiveFluorene-BODIPY copolymer synthesized by using the BODIPY monomer 2a inFIG. 12 as narrow-band monomer and fluorene as a general monomer. FIG.29B shows the emission spectra of the Fluorene-BODIPY Pdots in water. Asindicated by the spectra, these pure Fluorene-BODIPY Pdots showbroad-band emission. FIG. 29C shows emission spectra of the blendedPdots prepared from PFBT and Fluorene-BODIPY. The blended Pdot emissionexhibits a FWHM of 54 nm, which is narrower as compared with the purePdots. FIG. 29D shows other copolymers and blends of polymers andcopolymers for forming chromophoric polymer dots. FIGS. 29E-29H show thecorresponding optical properties and fluorescence emission spectra forthe different chromophoric polymer dots formed using the copolymers orblends of polymers and copolymers shown in FIG. 29D. FIG. 29E showsfluorescence spectra of chromophoric polymer dots of polymer 2b (singlepolymer) (I in FIG. 29D). FIG. 29F shows fluorescence spectra of blendedchromophoric polymer dots formed from II in FIG. 29D (10% mol BODIPYpolymer:PFBT=1:10). FIG. 29G shows fluorescence spectra of chromophoricpolymer dots of polymer 3b (single polymer) (III in FIG. 29D). FIG. 29Hshows fluorescence spectra of blended chromophoric polymer dots formedfrom IV in FIG. 29D (10% mol deep red BODIPYpolymer:PFTBT:PFBT=10:30:60).

FIG. 30A provides an example scheme showing the synthetic procedure forsynthesizing PFPPyBPh. FIG. 30B shows the UV-Vis and fluorescencespectra of PFPPyBPh in THF. FIG. 30C shows the UV-Vis and fluorescencespectra of PFPPyBPh chromophoric polymer dot.

FIG. 31A shows the schematic structures of narrow-band emissive polymerschemically cross-linked with two or more broad-band polymers.

FIG. 31B show an example of a narrow emissive polymer chemicallycross-linked with broad-band semiconducting polymer and the formation ofPdot-bioconjugates for specific cellular targeting. Two broad-bandfluorescent polymers containing benzothiadiazole (BT) and4,7-dithiophenyl-2,1,3-benzothiadiazole (TBT) are energy donors and thenarrow-band emissive polymer containing BODIPY is energy acceptor. Thethree fluorescent polymers with amine group can react with amphiphilicpolymer such as poly(styrene-co-maleic anhydride) (PSMA). Aftercross-linking reaction and Pdot formation, multi-step energy-transferinside Pdots results in narrow-band emissions.

FIG. 32 shows an example multi-step synthesis of BODIPY monomer 4a inFIG. 12 and copolymer that include a general monomer fluorene and BODIPYmonomer 4a with amine group via Yamamoto polymerization(PF5%540BODIPY4NH₂).

FIG. 33 shows an example synthesis of PF10BT polymer with an amine groupvia Suzuki coupling polymerization (PF10BT4NH₂).

FIG. 34 shows an example synthesis of copolymer that comprises a generalmonomer fluorene and BODIPY monomer 2a in FIG. 12 with an amine groupvia Yamamoto polymerization (PF5%540BODIPY4NH₂).

FIG. 35 shows an example synthesis of PFTBT copolymer with an aminegroup via Suzuki coupling polymerization (PF5TBT4NH₂).

FIG. 36 shows an example synthesis of copolymer that comprises a generalmonomer fluorene and BODIPY Monomer 3a in FIG. 12 with amine group viaYamamoto polymerization (PF5%680BODIPY4NH₂).

FIG. 37 shows an example multi-step synthesis of BODIPY monomer 5a inFIG. 12 and copolymer that includes a general monomer fluorene andBODIPY Monomer 5a with amine group via Yamamoto polymerization(PF5%570BODIPY4NH₂).

FIG. 38 shows an example multi-step synthesis of BODIPY monomer 8a inFIG. 12 and copolymer that comprises a general monomer fluorene andBODIPY Monomer 8a with amine group via Yamamoto polymerization(PF5%670BODIPY4NH₂).

FIG. 39 shows an example multi-step synthesis of BODIPY monomer 6a inFIG. 12 and copolymer that comprise a general monomer fluorene andBODIPY monomer 6a with amine group via Yamamoto polymerization(PF5%600BODIPY4NH₂).

FIG. 40 shows an example synthesis of copolymer that comprises2,1,3-benzoselenadiazole and a general monomer fluorene with amine groupvia Suzuki polymerization (PF47BSeD3NH₂).

FIG. 41A shows absorption and emission spectra of example blended Pdotsprepared from PF10BT4NH₂ and 540BODIPY fluorine copolymer with aminegroup and amphiphilic polymer PS-PEG-COOH. The blended Pdot emissionexhibits a FWHM of 39 nm without fluorene's emission under the 405 laserexcitation when blending ratio of 540BODIPY copolymer to PF10BT4NH₂ wasgreater than 30:70 and less than 70:30. FIG. 41B shows absorption andemission spectra of the blended Pdots prepared from PF46BT4NH₂ and570BODIPY fluorene copolymer with amine group and amphiphilic polymerPS-PEG-COOH. The blended Pdot emission exhibits a FWHM of 38 nm when ablending ratio of 570BODIPY copolymer to PF46BT4NH₂ was 30:70. FIG. 41Cshows absorption and emission spectra of the blended Pdots prepared fromPF46BT4NH₂ and 590BODIPY fluorene copolymer with amine group andamphiphilic polymer PS-PEG-COOH. The blended Pdot emission exhibits aFWHM of 55 nm when blending ratio of 590BODIPY copolymer to PF10BT4NH₂was greater than 35:65. FIG. 41D shows absorption and emission spectraof the blended Pdots prepared from PF47BSeD3NH₂ and 600BODIPY fluorenecopolymer with amine group and amphiphilic polymer PS-PEG-COOH. Theblended Pdot emission exhibits a FWHM of 38 nm when blending ratio of600BODIPY copolymer to PF47BSeD3NH₂ was greater than 30:70. FIG. 41Eshows absorption and emission spectra of the blended Pdots prepared fromPF46BT4NH₂, PF5TBT4NH₂ and 655BODIPY fluorene copolymer with amine groupand amphiphilic polymer PS-PEG-COOH. The blended Pdot emission exhibitsa FWHM of 38 nm when blending ratio of 600BODIPY copolymer to PF5TBT4NH₂and PF46BT4NH₂ was 20:10:70. FIG. 41F shows absorption and emissionspectra of the blended Pdots prepared from PF46BT4NH₂, PF5TBT4NH₂ and680BODIPY fluorene copolymer with amine group and amphiphilic polymerPS-PEG-COOH. The blended Pdot emission exhibits a FWHM of 44 nm whenblending ratio of 680BODIPY copolymer to PF5TBT4NH₂ and PF46BT4NH₂ was35:20:45.

FIG. 42A shows absorption and emission spectra of example cross-linked540Pdots prepared from PF10BT4NH₂ and 540BODIPY fluorene copolymer withamine group and reactive amphiphilic polymer PSMA; the cross-linked 590Pdots prepared from PF46BT4NH₂ and 590BODIPY fluorene copolymer withamine group and reactive amphiphilic polymer PSMA; the cross-linked 680Pdots prepared from PF46BT4NH₂, PF5TBT4NH₂ and 680BODIPY fluorenecopolymer with amine group and reactive amphiphilic polymer PSMA,respectively. The cross-linked 540, 590 and 680 Pdots emission exhibit aFWHM of 39 nm, 55 nm, and 44 nm, respectively. FIG. 42B shows absorptionand emission spectra of the cross-linked 570 Pdots prepared fromPF46BT4NH₂ and 570BODIPY fluorene copolymer with amine group andreactive amphiphilic polymer PSMA. The cross-linked Pdot emissionexhibits a FWHM of 37 nm when reactant ratio of 570BODIPY copolymer toPF46BT4NH₂ was 35:65. FIG. 42C shows absorption and emission spectra ofthe cross-linked 600 Pdots prepared from PF47BSeD3NH₂ and 600BODIPYfluorene copolymer with amine group and reactive amphiphilic polymerPSMA. The cross-linked Pdot emission exhibits a FWHM of 38 nm whenreactant ratio of 600BODIPY copolymer to PF47BSeD3NH₂ was 40:60. FIG.42D shows absorption and emission spectra of the cross-linked 655 Pdotsprepared from PF46BT4NH₂, PF5TBT4NH₂ and 655BODIPY fluorene copolymerwith amine group and reactive amphiphilic polymer PSMA. The cross-linkedPdot emission exhibits a FWHM of 36 nm when reactant ratio of 655BODIPYcopolymer to PF5TBT4NH₂ and PF46BT4NH₂ was 20:10:70.

FIG. 43 shows the synthesis of a narrow-band BODIPY monomer, a generalfluorene-vinyl-benzothiadiazol monomer FVBT, and the copolymerPFVBT-BODIPY. The absorption peak of the polymer is tuned to 492 nm tomatch the 488 nm laser generally used in biological applications.

FIG. 44A shows the UV-Vis absorption and fluorescence spectra ofPFVBT-BODIPY Pdots functionalized with PS-PEG-COOH and PSMA polymers.FIG. 44B show the comparison on the absorption profile between PFBT andPFVBT Pdots. FIG. 44C shows the optical performance of PFVBT-BODIPY2Pdots functionalized with PSPEG and PSMA polymers.

FIGS. 45A-45C shows a non-limiting list of examples of BODIPY basednarrow-band emissive polymers. Their absorption peak can be tuned fromthe visible to near infra-red region.

FIG. 46 shows chemical structures of three general broad-band polymersP1, P2, P3, and one narrow-band emissive polymer P4. The blended polymerdots made from the four polymers exhibit narrow-band emissions in thenear infrared region.

FIGS. 47A-47D show the synthetic procedure of the four polymers shown inFIG. 46.

FIG. 48 top panel shows the TEM images and size distribution histogrammeasured by DLS of the four-component Pdots (4-NIR Pdots) made from P1,P2, P3, P4 polymers. The bottom panel shows the TEM images and sizedistribution histogram measured by DLS of the three-component Pdots(3-NIR Pdots) made from P2, P3, and P4 polymers.

FIG. 49 shows the absorption spectra of 4-NIR Pdots (solid line), 3-NIRPdots (dashed line) and fluorescence spectra of 4-NIR Pdots (dot line,λ_(ex)=380 nm) and 3-NIR Pdots (dashed-dot-dot line, λ_(ex)=450 nm).

FIGS. 50A and B show single-particle fluorescence images of Qdot705 (a)and 3-NIR Pdots (b) obtained under identical excitation and detectionconditions. FIGS. 50C-D show the histograms of intensity distribution ofsingle-particle fluorescence for Qdot705 (mean=1200 CCD counts) and3-NIR Pdots (mean=3900 CCD counts).

FIGS. 51A and B show flow cytometry measurements of the intensitydistributions of MCF-7 cells labeled with 4-NIR Pdot-streptavidin (a,negative labeling, dot line; positive labeling, solid line) and 3-NIRPdot-streptiavidin. (b, negative labeling, dot line; positive labeling,solid line). All the negative and positive labelings were completed andmeasured under identical experimental conditions, only in the negativelabeling primary biotinylated antibody was absent.

FIGS. 52A and B show fluorescence images of MCF-7 cells labeled with3-NIR-SA probes. (a) Positive labeling using 3-NIR-Pdot-SA probe. (b)Negative labeling carried out under the same condition as (a) but in theabsence of biotinylated antibody on the surface of the MCF-7 cells. Fromleft to right: blue fluorescence from the nuclear stain Hoechst 34580;red fluorescence images from 3-NIR-SA probes; Nomarski (DIC) images; andcombined fluorescence images. Scale bars: 20 μm.

FIG. 53 shows chemical structures of a general emissive polymer PVK,europium complexes (Eu15F and EuDNM), a general non-emissive polymer PS,and a functional polymer PS-PEG-COOH.

FIG. 54 shows schematic illustration of a non-limiting example forforming Pdots using a general emissive PVK polymer and Eu complexes.

FIG. 55 shows spectroscopic and TEM characterizations of Eu15F/PolymerPdots. (a) Absorption spectra of Eu15F/PS nanoparticles, Eu15F/PVK Pdotsand pure PVK Pdots, and the emission spectra of PVK Pdots. (b) TEM imageof Eu15F/PVK Pdots. (c) Quantum yields of Eu15F/PS Pdots and Eu15F/PVKPdots versus the ratios of Eu15F.

FIG. 56 shows emission spectra of Eu15F/PVK Pdots with varying ratios ofEu15F.

FIG. 57 shows emission intensity comparison between Eu15F/PSnanoparticles and Eu15F/PVK Pdots with varying Eu15F ratios.

FIGS. 58A-C show spectroscopic and TEM characterizations ofEuDNM/Polymer Pdots. (a) Absorption spectra of EuDNM/PS nanoparticles,EuDNM/PVK Pdots and pure PVK Pdots, and the emission spectra of PVKPdots. (b) TEM image of EuDNM/PVK Pdots. (c) The quantum yields ofEuDNM/PS Pdots and Eu15F/PVK Pdots versus the ratios of EuDNM.

FIG. 59 shows emission intensity comparison between EuDNM/PSnanoparticles and EuDNM/PVK Pdots with varying ratios of EuDNM.

FIG. 60 shows bioconjugation scheme and the flow cytometry results ofthe two types of Eu/PVK Pdots.

FIG. 61 shows time-gated and un-gated fluorescence images of Eu15F/PVKPdots (left panel) and commercial R300 red fluorescence emittingnanoparticles (right panel).

FIG. 62 shows the distinguishing of Eu15F/PVK Pdots and commercial 300red fluorescence emitting nanoparticles based on their fluorescencelifetime difference. A shows the emission spectra of the Eu15F/PVK Pdotsand commercial R300 red fluorescence emitting nanoparticles. Theun-gated normal fluorescence image (left bottom; B) andtime-gated-fluorescence image (C) of Eu15F/PVK Pdots and commercial R300red fluorescence emitting nanoparticle mixture deposited on thecoverslip.

FIG. 63 shows time-gated fluorescence images of Eu15F/PVK Pdotsendocytosed by MCF-7 cells.

DETAILED DESCRIPTION OF THE INVENTION

Embodiments of the present invention relate to a novel class offluorescent probe, referred to as narrow-band emissive chromophoricpolymer dots, and their biomolecular conjugates for a variety ofapplications, including but not limited to flow cytometry, fluorescenceactivated sorting, immunofluorescence, immunohistochemistry,fluorescence multiplexing, single molecule imaging, single particletracking, protein folding, protein rotational dynamics, DNA and geneanalysis, protein analysis, metabolite analysis, lipid analysis, FRETbased sensors, high throughput screening, cell detection, bacteriadetection, virus detection, biomarker detection, cellular imaging, invivo imaging, bioorthogonal labeling, click reactions,fluorescence-based biological assays such as immunoassays andenzyme-based assays, and a variety of fluorescence techniques inbiological assays and measurements.

While not limited to any particular theory or concept, the presentinvention is based at least in-part on the fact that fluorescent Pdotsbased on semiconducting polymers typically possess broad emissionspectra with FWHM larger than 70 nm. Such a broad-band emission can be asignificant drawback for fluorescence techniques in biology. To overcomethis challenge with the current Pdots, the present invention providescompositions and methods to obtain next-generation Pdots withnarrow-band emissions. Furthermore, the present invention providescompositions and methods that allow bioconjugation to polymer dots whilealso maintaining their narrow-band emissions.

In some aspects, the properties of the narrow-band emissive polymers andpolymer dots can be dependent on the polymer structures. Therefore, thepolymer backbone (main chain), side chains, terminal units, andsubstituted groups can be varied to obtain specific properties. In someembodiments, the optical properties of the narrow-band polymer andpolymer dots can be tuned by varying the structures of the polymerbackbone (main chain). For example, the absorption and fluorescenceemission can be red-shifted by increasing the conjugation length of thepolymer backbone, or the absorption and fluorescence emission can beblue-shifted by decreasing the conjugation length of the polymerbackbone. As another example, the inclusion of BT monomer can increasethe photostability of the resulting polymer dot compared with polymersthat do not have BT in their polymer backbone.

In some embodiments, the optical properties of the narrow-band emissivepolymer and polymer dots can be modified by varying the side chains,terminal units, and substituent groups. For example, the fluorescenceemission wavelength can be tuned by attaching chromophoric units to theside-chains and/or termini. The emission bandwidth, fluorescence quantumyield, fluorescence lifetime, photostability, and other properties canalso be modified by varying the polymer side-chain and/or terminal unitsin addition to the polymer backbone. For example, fluorescence quantumyield can be increased in some polymer dots by attaching bulkyside-chain groups to minimize inter-chain interactions in polymer dots.In another example, the attachment and presence of anti-fade agents,such as derivatives of butylated hydroxytoluene, trolox, carotenoids,ascorbate, reduced glutathione, propyl gallate, propionic acid stearylester, hydroxyquinone, p-phenylenediamine, triphenylamine, betamercaptoethanol, trans-stilbene, imidazole, Mowiol, or combinationsthereof, or any other combinations of anti-fade agents known in the art,to the polymer via side chains, terminal units, backbone, and/orsubstituent groups, can increase quantum yield, photostability, or both.These anti-fade agents generally act as anti-oxidants to reduce oxygen,and/or act as scavengers of reactive oxygen species, and/or act tosuppress photogenerated hole polarons within the polymer dot. In apreferred embodiment, the anti-fade agent is hydrophobic in nature so asnot to adversely affect the packing and/or colloidal stability of thepolymer dot. In some embodiments, the absorption, emission peak,emission bandwidth, fluorescence quantum yield, fluorescence lifetime,photostability, and other properties of the narrow-band emissive polymerand polymer dots can also be modified by substituent groups on thepolymers. For example, the degree of electron-donating orelectron-withdrawing capability of the substituent groups can be used totune the optical properties. For example, the two-photon absorptioncross sections can be increased by modular structures such asdonor-pi-donor or donor-acceptor-donor units.

In some embodiments, the colloidal properties of the polymer dots can beimproved by varying the polymer backbone (main chain), side chains,terminal units, and substitutent groups. In some embodiments, thepolymer dots can include hydrophobic functional groups in theside-chains, terminal units, and/or substitutent groups. In otherembodiments, the polymer dots can include hydrophilic functional groupsin the side-chains, terminal units, and/or substitutent groups. Thelength, size, and nature of the hydrophobic/hydrophilic side chains canmodify the chain-chain interactions, and control the packing of thepolymers, and affect the colloidal stability and size of the polymerdots. The length, size, and nature of the hydrophobic/hydrophilic sidechains can also affect the absorption, emission peak, emissionbandwidth, fluorescence quantum yield, fluorescence lifetime,photostability, and other properties of the narrow-band emissive polymerand polymer dots. For example, a large number of very hydrophilicfunctional groups can reduced the brightness of the polymer dots, and/orbroaden the emission spectrum, and/or also adversely affect theircolloidal stability and non-specific binding properties.

Definitions

As used herein, the term “chromophoric polymer nanoparticle” or“chromophoric polymer dot” refers to a structure comprising one or morepolymers (e.g., chromophoric polymers) that have been formed into astable sub-micron sized particle. The chromophoric polymer nanoparticlesor chromophoric polymer dots of the present invention can, e.g., includea single polymer or a plurality of polymers that can be, e.g.,chemically crosslinked and/or physically blended. “Polymer dot” and“Pdot” can be used interchangeably to represent “chromophoricnanoparticle” or “chromophoric polymer dot”. The chromophoric polymerdots provided herein may be formed by any method known in the art,including without limitation, methods relying on precipitation, methodsrelying on the formation of emulsions (e.g. mini or micro emulsion), andmethods relying on condensation. Pdots described herein are differentand distinct from nanoparticles formed from an aggregate ofpolyelectrolytes.

As used herein, “polymer” is a molecule composed of at least 2 repeatingstructural units typically connected by covalent chemical bonds. Therepeating structural unit may be one type of monomer, and the resultingpolymer is a homopolymer. In some embodiments, the polymers can includetwo different types of monomers, or three different types of monomers,or more types of monomers. One of ordinary skill in the art willappreciate that the different types of monomers can be distributed alonga polymer chain in a variety of ways. For example, three different typesof monomers can be randomly distributed along the polymer. It willsimilarly be appreciated that the distribution of monomers along thepolymer can be represented in different ways. The number of repeatingstructural units (e.g., monomers) along the length of a polymer can berepresented by “n.” In some embodiments, n can range, e.g., from atleast 2, from at least 100, from at least 500, from at least 1000, fromat least 5000, or from at least 10,000, or from at least 100,000, orhigher. In certain embodiments, n can range from 2 to 10000, from 20 to10000, from 20 to 500, from 50 to 300, from 100 to 1000, or from 500 to10,000.

Polymers generally have extended molecular structures comprisingbackbones that optionally contain pendant side groups. The polymersprovided herein can include, but are not limited to, linear polymers andbranched polymers such as star polymers, comb polymers, brush polymers,ladders, and dendrimers. As described further herein, the polymers caninclude semiconducting polymers generally well known in the art.

As used herein, the term “chromophoric polymer” is a polymer in which atleast a portion of the polymer includes chromophoric units. The term“chromophore” is given its ordinary meaning in the art. A chromophoreabsorbs certain wavelength of light from UV to near infrared region, andmay be or may not be emissive. The chromophoric polymer can, e.g., be a“conjugated polymer”. The term “conjugated polymer” is recognized in theart. Electrons, holes, or electronic energy, can be conducted along theconjugated structure. In some embodiments, a large portion of thepolymer backbone can be conjugated. In some embodiments, the entirepolymer backbone can be conjugated. In some embodiments, the polymer caninclude conjugated structures in their side chains or termini. In someembodiments, the conjugated polymer can have conducting properties, e.g.the polymer can conduct electricity. In some embodiments, the conjugatedpolymer can have semiconducting properties, e.g. the polymers canexhibit a direct band gap, leading to an efficient absorption oremission at the band edge.

A “chromophoric unit” in this invention includes, but is not limited to,a unit of structures with delocalized pi-electrons, a unit of smallorganic dye molecules, and/or a unit of metal complexes. Examples ofchromophoric polymers can include polymers comprising units ofstructures with delocalized pi-electrons such as semiconductingpolymers, polymers comprising units of small organic dye molecules,polymers comprising units of metal complexes, and polymers comprisingunits of any combinations thereof. The chromophoric unit can beincorporated into the polymer backbone. The chromophoric unit can alsobe covalently attached to the side chain, or the terminal unit of thepolymer.

An “emission spectrum” of a polymer dot is defined as the spectrum ofwavelengths (or frequencies) of electromagnetic radiation emitted by thepolymer dot when it is excited to a higher energy state and thenreturned to a lower energy state. The width of the emission spectrum canbe characterized by its full width at half maximum (FWHM). The FWHM ofan emission spectrum is defined as the distance between points on theemission curve at which the emission intensity reaches half its maximumvalue. The emission properties of a polymer dot can also becharacterized by fluorescence quantum yield and fluorescence lifetime.The fluorescence quantum yield gives the efficiency of the fluorescenceprocess. It is defined as the ratio of the number of photons emitted tothe number of photons absorbed by the Pdots. The fluorescence lifetimeis defined as the average time the polymer dot stays in its excitedstate before emitting a photon. All the above defined parameters, suchas emission spectrum, FWHM, fluorescence quantum yield, and fluorescencelifetime can be experimentally measured. In this invention, theseparameters can be specifically used to characterize the narrow-bandemissive Pdots.

As used herein, the term “alkyl” refers to a straight or branched,saturated, aliphatic radical having the number of carbon atomsindicated. For example, C₁-C₆ alkyl includes, but is not limited to,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,tert-butyl, pentyl, isopentyl, hexyl, etc. Other alkyl groups include,but are not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl caninclude any number of carbons, such as 1-2, 1-3, 1-4, 1-5, 1-6, 1-7,1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 3-4, 3-5, 3-6, 4-5, 4-6 and 5-6. Thealkyl group is typically monovalent, but can be divalent, such as whenthe alkyl group links two moieties together. As used herein, the term“heteroalkyl” refers to a straight or branched, saturated, aliphaticradical of carbon atoms, where at least one of the carbon atoms isreplaced with a heteroatom, such as N, O or S. Additional heteroatomscan also be useful, including, but not limited to, B, Al, Si and P.

The term “lower” referred to above and hereinafter in connection withorganic radicals or compounds respectively defines a compound or radicalwhich can be branched or unbranched with up to and including 7,preferably up to and including 4 and (as unbranched) one or two carbonatoms.

As used herein, the term “alkylene” refers to an alkyl group, as definedabove, linking at least two other groups, i.e., a divalent hydrocarbonradical. The two moieties linked to the alkylene can be linked to thesame atom or different atoms of the alkylene. For instance, a straightchain alkylene can be the bivalent radical of —(CH₂)_(n), where n is 1,2, 3, 4, 5 or 6. Alkylene groups include, but are not limited to,methylene, ethylene, propylene, isopropylene, butylene, isobutylene,sec-butylene, pentylene and hexylene.

The groups described herein can be substituted or unsubstituted.Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be a variety of groups, such as alkyl, aryl,cyano (CN), amino, sulfide, aldehyde, ester, ether, acid, hydroxyl orhalide. Substituents can be a reactive group, such as but not limited tochloro, bromo, iodo, hydroxyl, or amino. Suitable substituents can beselected from: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′, -halogen,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′ C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —CN and —NO₂ in a numberranging from zero to (2m′+1), where m′ is the total number of carbonatoms in such radical. R′, R″ and R′″ each independently refer tohydrogen, unsubstituted (C₁-C₈)alkyl and heteroalkyl, unsubstitutedaryl, alkoxy or thioalkoxy groups, or aryl-(C₁-C₄)alkyl groups. When R′and R″ are attached to the same nitrogen atom, they can be combined withthe nitrogen atom to form a 5-, 6-, or 7-membered ring. For example,—NR′R″ is meant to include 1-pyrrolidinyl and 4-morpholinyl. From theabove discussion of substituents, one of skill in the art willunderstand that the term “alkyl” is meant to include groups such ashaloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g., —C(O)CH₃, —C(O)CF₃,—C(O)CH₂OCH₃, and the like).

As used herein, the term “alkoxy” refers to an alkyl group having anoxygen atom that either connects the alkoxy group to the point ofattachment or is linked to two carbons of the alkoxy group. Alkoxygroups include, for example, methoxy, ethoxy, propoxy, iso-propoxy,butoxy, 2-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, pentoxy, hexoxy,etc. The alkoxy groups can be further substituted with a variety ofsubstituents described within. For example, the alkoxy groups can besubstituted with halogens to form a “halo-alkoxy” group.

As used herein, the term “alkenyl” refers to either a straight chain orbranched hydrocarbon of 2 to 6 carbon atoms, having at least one doublebond. Examples of alkenyl groups include, but are not limited to, vinyl,propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl,1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl,1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl,1,5-hexadienyl, 2,4-hexadienyl, or 1,3,5-hexatrienyl.

As used herein, the term “alkenylene” refers to an alkenyl group, asdefined above, linking at least two other groups, i.e., a divalenthydrocarbon radical. The two moieties linked to the alkenylene can belinked to the same atom or different atoms of the alkenylene. Alkenylenegroups include, but are not limited to, ethenylene, propenylene,isopropenylene, butenylene, isobutenylene, sec-butenylene, pentenyleneand hexenylene.

As used herein, the term “alkynyl” refers to either a straight chain orbranched hydrocarbon of 2 to 6 carbon atoms, having at least one triplebond. Examples of alkynyl groups include, but are not limited to,acetylenyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl,butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl,1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl,1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl.

As used herein, the term “alkynylene” refers to an alkynyl group, asdefined above, linking at least two other groups, i.e., a divalenthydrocarbon radical. The two moieties linked to the alkynylene can belinked to the same atom or different atoms of the alkynylene. Alkynylenegroups include, but are not limited to, ethynylene, propynylene,isopropynylene, butynylene, sec-butynylene, pentynylene and hexynylene.

As used herein, the term “alkyl amine” refers to an alkyl groups asdefined within, having one or more amino groups. The amino groups can beprimary, secondary or tertiary. The alkyl amine can be furthersubstituted with a hydroxy group. Alkyl amines can include, but are notlimited to, ethyl amine, propyl amine, isopropyl amine, ethylene diamineand ethanolamine. The amino group can link the alkyl amine to the pointof attachment with the rest of the compound, be at the omega position ofthe alkyl group, or link together at least two carbon atoms of the alkylgroup.

As used herein, the term “halogen” or “halide” refers to fluorine,chlorine, bromine and iodine. As used herein, the term “haloalkyl”refers to alkyl as defined above where some or all of the hydrogen atomsare substituted with halogen atoms. Halogen (halo) preferably representschloro or fluoro, but may also be bromo or iodo. As used herein, theterm “halo-alkoxy” refers to an alkoxy group having at least onehalogen. Halo-alkoxy is as defined for alkoxy where some or all of thehydrogen atoms are substituted with halogen atoms. The alkoxy groups canbe substituted with 1, 2, 3, or more halogens. When all the hydrogensare replaced with a halogen, for example by fluorine, the compounds areper-substituted, for example, perfluorinated. Halo-alkoxy includes, butis not limited to, trifluoromethoxy, 2,2,2,-trifluoroethoxy,perfluoroethoxy, etc.

As used herein, the term “cycloalkyl” refers to a saturated or partiallyunsaturated, monocyclic, fused bicyclic or bridged polycyclic ringassembly containing from 3 to 12 ring atoms, or the number of atomsindicated. Monocyclic rings include, for example, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Bicyclic andpolycyclic rings include, for example, norbornane, decahydronaphthaleneand adamantane. For example, C₃₋₈cycloalkyl includes cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, and norbornane.

As used herein, the term “cycloalkylene” refers to a cycloalkyl group,as defined above, linking at least two other groups, i.e., a divalenthydrocarbon radical. The two moieties linked to the cycloalkylene can belinked to the same atom or different atoms of the cycloalkylene.Cycloalkylene groups include, but are not limited to, cyclopropylene,cyclobutylene, cyclopentylene, cyclohexylene, and cyclooctylene.

As used herein, the term “heterocycloalkyl” refers to a ring systemhaving from 3 ring members to about 20 ring members and from 1 to about5 heteroatoms such as N, O and S. Additional heteroatoms can also beuseful, including, but not limited to, B, Al, Si and P. The heteroatomscan also be oxidized, such as, but not limited to, —S(O)— and —S(O)₂—.

As used herein, the term “heterocycloalkylene” refers to aheterocycloalkyl group, as defined above, linking at least two othergroups. The two moieties linked to the heterocycloalkylene can be linkedto the same atom or different atoms of the heterocycloalkylene.

As used herein, the term “aryl” refers to a monocyclic or fusedbicyclic, tricyclic or greater, aromatic ring assembly containing 6 to16 ring carbon atoms. For example, aryl may be phenyl, benzyl, azulenylor naphthyl. “Arylene” means a divalent radical derived from an arylgroup. Aryl groups can be mono-, di- or tri-substituted by one, two orthree radicals selected from alkyl, alkoxy, aryl, hydroxy, halogen,cyano, amino, amino-alkyl, trifluoromethyl, alkylenedioxy andoxy-C₂-C₃-alkylene; all of which are optionally further substituted, forinstance as hereinbefore defined; or 1- or 2-naphthyl; or 1- or2-phenanthrenyl. Alkylenedioxy is a divalent substitute attached to twoadjacent carbon atoms of phenyl, e.g. methylenedioxy or ethylenedioxy.Oxy-C₂-C₃-alkylene is also a divalent substituent attached to twoadjacent carbon atoms of phenyl, e.g. oxyethylene or oxypropylene. Anexample for oxy-C₂-C₃-alkylene-phenyl is 2,3-dihydrobenzofuran-5-yl.

Aryl groups can include, but are not limited to, naphthyl, phenyl orphenyl mono- or disubstituted by alkoxy, phenyl, halogen, alkyl ortrifluoromethyl, phenyl or phenyl-mono- or disubstituted by alkoxy,halogen or trifluoromethyl, and in particular phenyl.

As used herein, the term “arylene” refers to an aryl group, as definedabove, linking at least two other groups. The two moieties linked to thearylene are linked to different atoms of the arylene. Arylene groupsinclude, but are not limited to, phenylene.

As used herein, the terms “alkoxy-aryl” or “aryloxy” refers to an arylgroup, as defined above, where one of the moieties linked to the aryl islinked through an oxygen atom. Alkoxy-aryl groups include, but are notlimited to, phenoxy (C₆H₅O—). The present invention also includesalkoxy-heteroaryl or heteroaryloxy groups.

As used herein, the term “heteroaryl” refers to a monocyclic or fusedbicyclic or tricyclic aromatic ring assembly containing 5 to 16 ringatoms, where from 1 to 4 of the ring atoms are a heteroatom each N, O orS. For example, heteroaryl includes pyridyl, indolyl, indazolyl,quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl,furanyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl,triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, or any otherradicals substituted, especially mono- or di-substituted, by e.g. alkyl,nitro or halogen. Suitable groups for the present invention can alsoinclude heteroarylene and heterarylene-oxy groups similar to thedescription above for arylene and arylene-oxy groups.

Similarly, aryl and heteroaryl groups described herein can besubstituted or unsubstituted. Substituents for the aryl and heteroarylgroups are varied, such as alkyl, aryl, CN, amino, sulfide, aldehyde,ester, ether, acid, hydroxyl or halide. Substituents can be a reactivegroup, such as but not limited to chloro, bromo, iodo, hydroxyl, oramino. Substituents can be selected from: -halogen, —OR′, —OC(O)R′,—NR′R″, —SR′, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O) R′, —OC(O)NR′R″,—NR″C(O)R′, —NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR ′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —N₃, —CH(Ph)₂, in anumber ranging from zero to the total number of open valences on thearomatic ring system; and where R′, R″ and R′″ are independentlyselected from hydrogen, (C₁-C₈)alkyl and heteroalkyl, unsubstituted aryland heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl, and (unsubstitutedaryl)oxy-(C₁-C₄)alkyl.

As used herein, the term “alkyl-aryl” refers to a radical having analkyl component and an aryl component, where the alkyl component linksthe aryl component to the point of attachment. The alkyl component is asdefined above, except that the alkyl component is at least divalent inorder to link to the aryl component and to the point of attachment. Insome instances, the alkyl component can be absent. The aryl component isas defined above. Examples of alkyl-aryl groups include, but are notlimited to, benzyl. The present invention also includes alkyl-heteroarylgroups.

As used herein, the term “alkenyl-aryl” refers to a radical having bothan alkenyl component and an aryl component, where the alkenyl componentlinks the aryl component to the point of attachment. The alkenylcomponent is as defined above, except that the alkenyl component is atleast divalent in order to link to the aryl component and to the pointof attachment. The aryl component is as defined above. Examples ofalkenyl-aryl include ethenyl-phenyl, among others. The present inventionalso includes alkenyl-heteroaryl groups.

As used herein, the term “alkynyl-aryl” refers to a radical having bothan alkynyl component and a aryl component, where the alkynyl componentlinks the aryl component to the point of attachment. The alkynylcomponent is as defined above, except that the alkynyl component is atleast divalent in order to link to the aryl component and to the pointof attachment. The aryl component is as defined above. Examples ofalkynyl-aryl include ethynyl-phenyl, among others. The present inventionalso includes alkynyl-heteroaryl groups.

As will be appreciated by one of ordinary skill in the art, the variouschemical terms defined herein can be used for describing chemicalstructures of the polymers and monomers of the present invention. Forexample, a variety of the monomer derivatives (e.g., BODIPY derivatives)can include a variety of the chemical substituents and groups describedherein. For example, in some embodiments, derivatives of the variousmonomers can be substituted with hydrogen, deuterium, alkyl, aralkyl,aryl, alkoxy-aryl, N-dialkyl-4-phenyl, N-diphenyl-4-phenyl,N-dialkoxyphenyl-4-phenyl, amino, sulfide, aldehyde, ester, ether, acid,and/or hydroxyl.

Optical Properties of Narrow-Band Emissive Chromophoric Polymer Dots

The present invention provides, in one embodiment, chromophoric polymerdots with narrow-band emissions. The emission wavelength of the polymerdots can vary from ultraviolet to the near infrared region. Thechromophoric polymer dot includes at least one chromophoric polymer. Asprovided herein, the chemical composition and structure of thechromophoric polymer can be tuned to obtain small bandwidth (FWHM) ofthe Pdot emission. Other species such as narrow-band emissive units,metal complexes or inorganic materials can be blended or chemicallycross linked within the chromophric polymer dots to obtain smallbandwidth (FWHM) of the Pdot emission. In some embodiments, the FWHM isless than about 70 nm. In some embodiments, the FWHM is less than about65 nm. In some embodiments, the FWHM is less than about 60 nm. In someembodiments, the FWHM is less than about 55 nm. In some embodiments, theFWHM is less than about 50 nm. In some embodiments, the FWHM is lessthan about 45 nm. In some embodiments, the FWHM is less than about 40nm. In some embodiments, the FWHM is less than about 35 nm. In someembodiments, the FWHM is less than about 30 nm. In some embodiments, theFWHM is less than about 25 nm. In certain embodiments, the FWHM is lessthan about 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18 nm, 17 nm, 16nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less. In someembodiments, the FWHM of the polymer dots described herein can rangebetween about 5 nm to about 70 nm, from about 10 nm to about 60 nm, fromabout 20 nm to about 50 nm, or from about 30 nm to about 45 nm.

In some embodiments, the chemical composition and structure of thechromophoric polymer in the polymer dots can affect the absorptionspectrum of the narrow-band emissive Pdots. The absorption peak canshift from ultra-violet region to near infrared region. In someembodiments, the absorption peak of the narrow-band emissive polymerdots can be tuned to a certain laser wavelength. In some embodiments,for example, the absorption peak can be tuned to around 266 nm. In someembodiments the absorption peak can be tuned to around 355 nm. In someembodiments, the absorption peak can be tuned to around 405 nm. In someembodiments, the absorption peak can be tuned to around 450 nm. In someembodiments, the absorption peak can be tuned to around 488 nm. In someembodiments, the absorption peak can be tuned to around 532 nm. In someembodiments, the absorption peak can be tuned to around 560 nm. In someembodiments, the absorption peak can be tuned to around 635 nm. In someembodiments, the absorption peak can be tuned to around 655 nm. In someembodiments, the absorption peak can be tuned to around 700 nm. In someembodiments, the absorption peak can be tuned to around 750 nm. In someembodiments, the absorption peak can be tuned to around 800 nm. In someembodiments, the absorption peak can be tuned to around 900 nm. In someembodiments, the absorption peak can be tuned to around 980 nm. In someembodiments, the absorption peak can be tuned to around 1064 nm.

In certain embodiments, the chemical composition and structure of thechromophoric polymer in the polymer dots can affect the fluorescencequantum yield of the narrow-band emissive Pdots. The fluorescencequantum yield, for example, can vary from 100% to 0.1%. In someembodiments, the quantum yield can be greater than about 90%. In someembodiments, the quantum yield can be greater than about 80%. In someembodiments, the quantum yield can be greater than about 70%. In someembodiments, the quantum yield can be greater than about 60%. In someembodiments, the quantum yield can be greater than about 50%. In someembodiments, the quantum yield can be greater than about 40%. In someembodiments, the quantum yield can be greater than about 30%. In someembodiments, the quantum yield can be greater than about 20%. In someembodiments, the quantum yield can be greater than about 10%. In someembodiments, the quantum yield can be greater than about 5%. In someembodiments, the quantum yield can be greater than about 1%.

In some embodiments, the narrow-band emissive Pdots have both narrowemission FWHM and high fluorescence quantum yield. For example, thenarrow-band Pdots can have an emission FWHM less than 70 nm andfluorescence quantum yield larger than 10%. The narrow-band Pdots canhave an emission FWHM less than 60 nm and fluorescence quantum yieldlarger than 10%. The narrow-band Pdots can have an emission FWHM lessthan 50 nm and fluorescence quantum yield larger than 10%. Thenarrow-band Pdots can have an emission FWHM less than 40 nm andfluorescence quantum yield larger than 10%. The narrow-band Pdots canhave an emission FWHM less than 30 nm and fluorescence quantum yieldlarger than 10%. The narrow-band Pdots can have an emission FWHM lessthan 20 nm and fluorescence quantum yield larger than 10%. In certainembodiments, the quantum yield is greater than 20%, greater than 30%,greater than 40%, greater than 50%, greater than 60%, greater than 70%,greater than 80%, or greater than 90%.

In some embodiments, the narrow-band emissive Pdots may have a secondaryemission peak. For example, when the narrow-band monomers as energyacceptors are copolymerized with other monomer donors to producenarrow-band emissive Pdots, the final Pdots may have secondary peakbecause of incomplete fluorescence quenching. In some embodiments, thenarrow-band emissive Pdots may also have a secondary peak in thecomposite Pdot chemically cross-linked with fluorescent dyes (e.g.,fluorescent polymers and/or fluorescent small molecules), metalcomplexes, lanthanide complexes, inorganic quantum dots etc. Besides thenarrow emission with FWHM less than 70 nm for the main peak, thesecondary peak in the Pdots is less than 30% of the maximum intensity ofthe main narrow-band emission. In some embodiments, the secondary peakin the Pdots is less than 25% of the maximum intensity of the mainnarrow-band emission. In some embodiments, the secondary peak in thePdots is less than 20% of the maximum intensity of the main narrow-bandemission. In some embodiments, the secondary peak in the Pdots is lessthan 10% of the maximum intensity of the main narrow-band emission. Insome embodiments, the secondary peak in the Pdots is less than 5% of themaximum intensity of the main narrow-band emission. In some embodiments,the secondary peak in the Pdots is less than 1% of the maximum intensityof the main narrow-band emission, or less. Also, in certain embodiments,the quantum yield is greater than 10%, greater than 20%, greater than30%, greater than 40%, greater than 50%, greater than 60%, greater than70%, greater than 80%, or greater than 90%.

In certain embodiments, the chemical composition and structure of thechromophoric polymer in the polymer dots can affect the fluorescencelifetime of the narrow-band emissive Pdots. The fluorescence lifetimecan vary from 10 ps to 1 ms. In some embodiments, the fluorescencelifetime varies from 10 ps to 100 ps. In some embodiments, thefluorescence lifetime varies from 100 ps to 1 ns. In some embodiments,the fluorescence lifetime varies from 1 ns to 10 ns. In someembodiments, the fluorescence lifetime varies from 10 ns to 100 ns. Insome embodiments, the fluorescence lifetime varies from 100 ns to 1 μs.In some embodiments, the fluorescence lifetime varies from 1 μs to 10μs. In some embodiments, the fluorescence lifetime varies from 10 μs to100 μs. In some embodiments, the fluorescence lifetime varies from 100μs to 1 ms.

In certain embodiments, the narrow-band emissive Pdots can becharacterized by their stability. The optical properties (e.g. emissionspectrum, emission band width, fluorescence quantum yield, fluorescencelifetime, side peaks, brightness at the particular wavelength oremission intensity at a particular wavelength) can be stable for over 1day, or 1 week, or 2 weeks, or 1 month, or 2 months, or 3 months, or 6months, or 1 year, or longer. The stable fluorescence quantum yieldmeans that the fluorescence quantum yield of the narrow-band emissiondoes not change by more than 5%, or 10%, or 20%, or 50%, or higher. Thestable emission spectrum means that intensity ratio of the secondarypeak relative to the main peak doesn't change by more than 5%, or 10%,or 20%, or 50%, or higher.

In some embodiments, the narrow-band emissive Pdots can possess all ofthe following characteristics: (1) Narrow band emission that has a FWHMof less than 70 nm, preferably less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm; (2) High quantum yieldthat is greater than 5%, preferably greater than 10%, preferably greaterthan 20%, greater than 30%, greater than 40%, greater than 50%, greaterthan 60%, greater than 70%, greater than 80%, or greater than 90%; (3)With a secondary emission peak that is less than 30% of the main peak,preferably less than 20%, less than 10%, less than 5%, or less than 1%;(4) Have high stability over at least 2 weeks, preferably 1 month, 2month, 3 month, 6 months, 1 year, or longer.

Compositions of Narrow-Band Emissive Chromophoric Polymer Dots

The present invention can include polymer dots, e.g., narrow-bandemissive chromophoric polymer dots. As described further herein, thepresent invention includes a wide variety of polymer dots that exhibitnarrow band emission properties (e.g., a FWHM less than 70 nm). Asdescribed further herein, the variety of polymer dots of the presentinvention can include polymers that have a narrow band emissive unit(e.g., a narrow band monomer and/or a narrow band unit). For example,the present invention can include a homopolymer or heteropolymerincluding a narrow band monomer, such as BODIPY and/or BODIPY derivativemonomer, a squaraine and/or squaraine derivative, a metal complex and/ormetal complex derivative monomer, a porphyrin and/or porphyrinderivative monomer, a phthalocyanine and/or phthalocynanine derivativemonomer, a lanthanide complex and/or lanthanide complex derivativemonomer, a perylene and/or perylene derivative monomer, a cyanine and/orcyanine derivative monomer, a rhodamine and/or rhodamine derivativemonomer, a coumarin and/or coumarin derivative monomer, and/or axanthene and/or xanthene derivative monomer. A narrow band unit can be,e.g., a narrow band monomer or a fluorescent nanoparticle embedded in orattached to the polymer dot. The fluorescent nanoparticle can be, e.g.,a quantum dot. A narrow band unit can also include a polymer orfluorescent dye molecule that gives a narrow emission in a polymer dotof the present invention.

The narrow band monomers can be integrated into a heteropolymer withother general monomers that can, e.g., act as energy donors. Forexample, the general monomers can include an emission spectrum that istuned to substantially overlap the absorption spectrum of a narrow bandmonomer, thereby acting as an energy donor for the narrow band monomer.The energy transfer, e.g., can occur along the backbone of a polymer(e.g., intrachain) or between multiple polymer backbones (e.g.,interchain). In some embodiments, the narrow band units (e.g., narrowband monomers) can be attached (e.g., covalently attached) to a polymerbackbone or sidechain of the polymer. For example, the narrow band unit(e.g., a narrow band monomer) can be attached to a general monomer thatcan include an emission spectrum that is tuned to substantially overlapthe absorption spectrum of a narrow band unit, thereby acting as anenergy donor for the narrow band unit. The general monomers can includea wide variety of structures that are further described herein (e.g.,D1, D2, D2′, monomers of P1-P10, and/or M1-M10). In some embodiments,the general monomers can include, e.g., fluorene, a fluorene derivative,a phenyl vinylene, a phenyl vinylene derivative, a phenylene, aphenylene derivative, a benzothiazole, a benzothiazole derivative, athiophene, a thiophene derivative, a carbazole fluorene, and/or acarbazole fluorene derivative. As also described herein, the variouspolymers used in the polymer dots can be combined in a variety of ways.For example, the polymers of the present invention can be chemicallycrosslinked and/or physically blended in the polymer dots. The polymersdescribed herein can further include at least one functional group for,e.g., conjugation reactions, such as for bioconjugation reactions toantibodies or other biomolecules further described herein. The presentinvention further includes compositions including the polymer dotsdescribed herein. The compositions of the present invention can include,e.g., polymer dots described herein suspended in a solvent (e.g., anaqueous solution).

In some embodiments, the narrow-band emissive chromophoric polymer dotsinclude at least one narrow-band emissive polymer. The narrow-bandemissive polymer can be a homopolymer or a heteropolymer (e.g., acopolymer). The narrow-band emissive polymers may have broad-bandemissions in good solvents. However, the final Pdots made from thenarrow-band polymers have narrow-band emissions. In certain embodiments,the chromophoric polymer dots can include luminescent semiconductingpolymer with delocalized pi-electrons. The term “semiconducting polymer”is recognized in the art. Conventional luminescent semiconductingpolymers include, but are not limited to fluorene polymers, phenylenevinylene polymers, phenylene polymers, benzothiadiazole polymers,thiophene polymers, carbazole polymers and related copolymers. Whilethose conventional semiconducting polymers typically have broad-bandemissions, narrow-band emissive polymers in this invention includechemical units such as narrow-band monomers so that the final Pdots givenarrow-band emissions. The emission FWHM of the final Pdots is less than70 nm. In certain embodiments, the FWHM is less than 60 nm, less than 50nm, less than 40 nm, less than 30 nm, or less than 20 nm. Thenarrow-band monomers include but are not limited to BODIPY, squaraine,porphyrin, metallophyrin, metal complexes, lanthanide complexes,phthalocyanine, perylene, rhodamine, coumarin, xanthene, cyanine, andtheir derivatives. A non-limiting list of the narrow-band emissivemonomers (or chemical units) and a non-limiting list of the narrow-bandemissive polymer can be, for example, found in the attached figures.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude narrow-band monomers. The narrow-band emissive polymer dots canalso include other monomers that are broad-band emissive. Thenarrow-band monomers can be energy acceptors and other monomers can beenergy donors. For example, polymer dots of the present invention caninclude condensed polymer nanoparticles that have intrachain energytransfer between, e.g., a narrow-band monomer and one or more generalmonomers on the same polymer chain. The polymer dots can also haveinterchain energy transfer in which a condensed polymer nanoparticle caninclude two or more polymer chains physically blended and/or chemicallycrosslinked together. For interchain energy transfer, one of the chainsmay include a narrow-band monomer and another chain may include one ormore general monomers that can act as an energy donor to the narrow bandmonomer, which is an energy acceptor. Some of the polymer dots caninclude both intrachain and interchain energy transfer. In someinstances, the combination of intrachain and interchain energy transfercan increase the quantum yield of the polymer dots. In certainembodiments, the final Pdots can exhibit narrow-band emissions becauseof energy transfer to the narrow-band monomers. The narrow-band emissivechromophoric polymers in good solvents may exhibit broad emissions ornarrow emissions. However, their nanoparticle form gives narrow-bandemissions. The emission FWHM of the above Pdots is less than 70 nm. Incertain embodiments, the FWHM is less than 60 nm, less than 50 nm, lessthan 40 nm, less than 30 nm, or less than 20 nm. In some embodiments,the FWHM of the polymer dots described herein can range between about 5nm to about 70 nm, from about 10 nm to about 60 nm, from about 20 nm toabout 50 nm, or from about 30 nm to about 45 nm. In some embodiments,the narrow-band emissive Pdots are narrow band emission without relyingon the formation of any defined secondary structures, such as betaphase.

In some embodiments, the narrow-band emissive polymer is a homopolymerthat includes only narrow-band emissive monomer (FIG. 1A). As describedfurther herein, example narrow band monomers can include BODIPY and/or aBODIPY derivative monomer, a squaraine and/or squaraine derivative, ametal complex and/or metal complex derivative monomer, a porphyrinand/or porphyrin derivative monomer, a phthalocyanine and/orphthalocynanine derivative monomer, a lanthanide complex and/orlanthanide complex derivative monomer, a perylene and/or perylenederivative monomer, a cyanine and/or cyanine derivative monomer, arhodamine and/or rhodamine derivative monomer, a coumarin and/orcoumarin derivative monomer, and/or a xanthene and/or xanthenederivative monomer. In some embodiments, the narrow-band emissivepolymer is a two-unit copolymer that includes one narrow-band monomerand one general monomer (e.g., D, D1, D2, and/or D2′) (FIG. 1B). In someembodiments, the general monomers can be broad-band emissive. Thegeneral monomer can be an energy donor and the narrow-band monomer canbe an energy-acceptor. Energy-transfer inside Pdots can result innarrow-band emissions. In some embodiments, the narrow-band emissivepolymer is a three-unit copolymer that includes one narrow-band monomerand two general monomers such as general monomer 1 and general monomer 2(e.g., selected from D, D1, D2, and/or D2′) (FIG. 1C). The narrow-bandmonomer can be an energy-acceptor, general monomer 1 can be anenergy-donor, general monomer 2 can also be a donor to the narrow-bandmonomer. In some embodiments, general monomer 2 can be anenergy-acceptor from general monomer 1 and simultaneously anenergy-donor to the narrow-band monomer. Both general monomer 1 andgeneral monomer 2 can be broad-band emissive. However, multi-stepenergy-transfer inside Pdots can result in narrow-band emissions. Incertain embodiments, the narrow-band emissive polymer can be aheteropolymer, such as a multi-unit (>3) copolymer, that includes atleast one type of narrow-band emissive monomer so that the final Pdotsgive narrow-band emissions.

In some embodiments, the narrow-band emissive polymer is copolymer thatincludes the narrow-band unit cross-linked with the side-chains (FIG.1D). The copolymer can include 2 types of general monomers, or 3 typesof general monomers, or more than 3 types of general monomers (e.g.,selected from D, D1, D2, and/or D2′). However, the narrow-band emissivepolymer can include at least one type of narrow-band emissive unit inthe side-chains. The copolymer backbone can be an energy-donor, and thenarrow-band emissive unit can be an energy-acceptor. Energy-transferinside Pdots results in narrow-band emissions. In some embodiments, thenarrow-band emissive polymer is a homopolymer that includes thenarrow-band unit cross-linked with the side-chains (FIG. 1E). Thehomopolymer backbone can be an energy-donor, and the narrow-band unitcan be an energy-acceptor. Energy-transfer inside Pdots can result innarrow-band emissions.

In some embodiments, the narrow-band emissive polymer can be a polymerthat includes a narrow-band monomer attached to one terminus or bothtermini of the linear polymer (FIG. 1F) or all termini in case of abranched polymer. The polymer can, e.g., include one type of a generalmonomer (e.g., any one of D, D1, D2, or D2′), or two types of generalmonomers (e.g., any one of D, D1, D2 or D2′), or three types of generalmonomers, or more than three types of general monomers. In someembodiments, the narrow-band emissive polymer can include at least onetype of narrow-band emissive unit in one terminus or both termini of thelinear polymer (e.g., FIG. 1F) or all termini in case of a branchedpolymer. The polymer backbone can be an energy-donor, and thenarrow-band emissive unit can be an energy-acceptor. Energy-transferinside Pdots results in narrow-band emissions. In some embodiments, thenarrow-band emissive polymer can be a homopolymer or heteropolymer thatincludes the narrow-band unit attached to the terminus of the polymer.The homopolymer or heteropolymer backbone can be an energy-donor, andthe narrow-band unit can be an energy-acceptor. Energy-transfer insidePdots can result in narrow-band emissions.

FIGS. 1G-L show other examples of schematic structures for thenarrow-band emissive polymers that can include, e.g., general monomersas donors (D) and narrow band monomers as acceptors (A). In someaspects, the donors can absorb energy and transfer the energy, eitherdirectly or indirectly (e.g. by cascade energy transfer), to thenarrow-band monomers. Besides the general monomer and narrow bandmonomer, these polymers can also include functional monomers (F) thatprovide reactive functional groups for, e.g., chemical reactions andbioconjugation reactions. The functional monomers can be copolymerizedwith the general monomers and narrow band monomers (e.g., FIG. 1G), orcross-linked with these two kinds of monomers. The functional monomerscan be used as a terminus (or for both termini) of the polymers (e.g.,FIG. 1H and FIG. 1K). Functional groups can be included either in thegeneral monomer or the narrow band monomers (e.g., FIG. 1I). In someembodiments, the narrow band monomers can also be copolymerized with anyof the general polymers to synthesize a narrow-band emissive copolymeror heteropolymer that contains more than two types of monomers (e.g.,FIG. 1J). The narrow band monomer can be covalently attached to theside-chains of the polymer (e.g, FIG. 1I). In some embodiments, thenarrow band emissive units can be covalently attached to the terminus ofthe polymer. In some embodiments, the narrow band emissive units can bephysically mixed or blended with conventional semiconducting polymers toform narrow-band emissive polymer dots. In one embodiment, the narrowband emissive units can be covalently cross-linked with conventionalsemiconducting polymers to form narrow-band emissive polymer dots. Theconventional semiconducting polymers can absorb energy and transfer theenergy, either directly or indirectly (e.g. by cascade energy transfer)to the narrow band monomer.

All the narrow-band emissive polymers described above in FIGS. 1A-L can,e.g., be physically blended or chemically cross-linked with one or moregeneral broad-band polymers. In some aspects, the broad-band polymerscan be energy donors and the narrow-band emissive polymer can be energyacceptors. Multi-step energy transfer can occur from the broad-bandpolymer to the narrow-band emissive polymer so that the polymer dotsgive narrow-band emissions. One example of chemical crosslinking isshown in FIG. 31A. The chemical cross-linking between polymers can usethe functional reactive groups such as haloformyl, hydroxyl, aldehyde,alkenyl, alkynyl, anhydride, carboxamide, amines, azo compound,carbonate, carboxylate, carboxyl, cyanates, ester, haloalkane, imine,isocyanates, nitrile, nitro, phosphino, phosphate, phosphate, pyridyl,sulfonyl, sulfonic acid, sulfoxide, thiol groups. These functionalgroups can be attached to the side chains and/or the terminus of eachpolymer chain.

As described herein, the present invention can include general monomersthat can be polymerized with the narrow band monomers disclosed herein.FIG. 2A provides a non-limited list of example general monomers (D). Insome embodiments, the general monomer can act as an energy donor for anarrow band monomer in the polymer. A variety of derivatized monomericunits can be used. For example, for the structures shown in FIG. 2A,each of R¹, R², R³ and R⁴ can be independently selected from, but arenot limited to, alkyl, phenyl, alkyl-substituted phenyl,alkyl-substituted fluorenyl and alkyl-substituted carbazolyl. Alkylsubstituted phenyl can include 2-alkylphenyl, 3-alkylphenyl,4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl, 7-triphenylaminyl-9,9-dialkyl-substitute fluorenyl and7-diphenylaminyl-9,9-dialkyl-substitute fluorenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃ wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. The general monomers can also besubstituted with other substituents as defined herein.

In certain embodiments, a polymer can include one or more types ofgeneral monomers. As shown in FIGS. 3A-E, three example types of generalmonomers are shown, D1, D2 and D2′. Each of the general D1 type monomerscan be copolymerized with each of the D2 and D2′ type monomers and onenarrow-band monomer to obtain narrow-band emissive polymer. Any of theD1 type monomers or D2 type monomers can also be separately used tocopolymerize with one narrow-band monomer to obtain the narrow-bandemissive polymers as, e.g., in FIGS. 1B and 1E. For the structures shownin FIG. 3A, a variety of substituents can be attached to the basestructures. For example, each of R¹, R², R³, R³, R⁴, R⁵, and R⁶ can beindependently selected from the group consisting of, but not limited to,hydrogen (H), deuterium (D), halogen, direct or branched alkyl,heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-) substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-) substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The alkyl substituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃ wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. The general monomers can also besubstituted with other substituents as defined herein. As shown in FIG.3A, each of X, X¹, and X² can be independently selected from the groupconsisting of carbon (C), silicon (Si), and germanium (Ge). Z, Z¹, Z²can be selected from the group consisting of oxygen (O), sulfur (S), andselenium (Se).

FIG. 3B shows a non-limiting list of general donors in the narrow-bandemissive polymers. As shown in the chemical structures of donors in FIG.3B, each of X, X¹, X², X³, X⁴, Q, Z, Z¹, and Z² can be heteroatoms,e.g., and can be independently selected from the group consisting of, O,S, Se, Te, N and so on. Each of R¹ and R² is independently selected fromnon-limiting examples of hydrogen (H), deuterium (D), halogen, direct orbranched alkyl, heteroalkyl, heterocycloalkyl, heterocycloalkylene,alkoxy, aryl, hydroxyl, cyano, nitro, ether and its derivatives, esterand its derivatives, alkyl ketone, alkylester, arylester, alkynyl, alkylamine, fluoroalkyl, fluoroaryl, and polyalkalene (e.g,mehtoxyethoxyethoxy, ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50),phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedphenyl, pyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted bipyridyl tripyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl,pyrrolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrrolyl, pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.

In some embodiments, the general donors can be selected (but not limitedto) from the group shown in FIGS. 3C, 3D and 3E. As shown in the variousD2 and D2′ structures in FIGS. 3C, 3D, and 3E, each of R¹, R², R³, andR⁴ can be independently selected from non-limiting examples of hydrogen(H), deuterium (D), halogen, direct or branched alkyl, heteroalkyl,heterocycloalkyl, heterocycloalkylene, alkoxy, aryl, hydroxyl, cyano,nitro, ether and its derivatives, ester and its derivatives, alkylketone, alkylester, arylester, alkynyl, alkyl amine, fluoroalkyl,fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy,and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted phenyl, pyridyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl, pyrazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyrazolyl,oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedoxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted thiazolyl, imidazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted imidazolyl, pyrazinyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrazinyl,benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude boron-dipyrromethene(4,4-difluoro-4-bora-3a,4a-diaza-s-indacene, BODIPY) and theirderivatives as narrow-band monomers. BODIPY monomers and theirderivatives include but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, BODIPY extended systems and other BODIPYderivatives. The narrow-band emissive polymers can also include anyother monomers. The BODIPY based-monomers can be energy acceptors andother monomers can be energy donors so that the final Pdots can exhibitnarrow-band emissions. The narrow-band emissive chromophoric polymers ingood solvents may exhibit broad emissions or narrow emissions. However,their nanoparticle form gives narrow-band emissions. The emission FWHMof the above Pdots is less than 70 nm. In certain embodiments, the FWHMis less than 60 nm, less than 50 nm, less than 40 nm, less than 30 nm,or less than 20 nm.

Suitable narrow band monomers of the present invention can includeBODIPY derivatives and other boron-containing monomers. FIGS. 2B-2Nshows a non-limiting list of examples of the narrow-band emissivecopolymer including one general monomer chosen from FIG. 2A anddifferent BODIPY derivatives or other boron-containing units asnarrow-band monomers. The narrow band monomers in FIGS. 2B-2L caninclude a variety of substituents defined herein. For example, for thestructures shown in FIGS. 2B and 2C, each of R¹, R², R⁴ and R⁵ can beindependently selected from the group consisting of fluorine (F),phenyl, naphthyl, alkyl-substituted phenyl, alkyl-substituted naphthyland alkyl-substituted thiophenyl. Alkyl substituted phenyl can include2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, and 3,4-dialkylphenyl. Alkyl-substituted naphthyl caninclude 3-alkyl-substituted naphthyl, 4-alkyl-substituted naphthyl,6-alkyl-substituted naphthyl and 7-alkyl-substituted naphthyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, 4-alkylthiophenyl. The alkyl substituents can includeC_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃ wherein nis 1 to 20. In some embodiments, n can be between 1 to 50 or higher. R³can be selected from the group consisting of phenyl, alkyl-substitutedphenyl, and alkyl-substituted thiophenyl, and in some cases, from cyano(CN), fluorine (F), and trifluoro (CF₃). Additional narrow-band monomersare shown in FIGS. 2D-2G in which each of R¹ and R² are independentlyselected from the group consisting of, but not limited to, fluorine,alkyl, phenyl, alkyl-substituted phenyl, alkyl-substituted fluorenyl andalkyl-substituted carbazolyl. Alkyl substituted phenyl can include2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, and 3,4-dialkylphenyl. Alkyl-substituted fluorenylcan include 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃ wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher.

FIGS. 2H and 2I show additional example monomers in which each of R¹,R², R³ and R⁴ are independently selected from the group consisting of,but not limited to, fluorine, alkyl, phenyl, alkyl-substituted phenyl,alkyl-substituted fluorenyl, alkyl-substituted carbazolyl andalkyl-substituted thiophenyl. Alkyl substituted phenyl can include2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, and 3,4-dialkylphenyl. Alkyl-substituted fluorenylcan include 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substitutedthiophenyl can include 3-alkyl-substituted thiophenyl,4-alkyl-substituted thiophenyl, 5-alkyl-substituted thiophenyl,3,4-dialkyl-substituted thiophenyl, 3,5-dialkyl-substituted thiophenyland 4,5-dialkyl-substituted thiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher.

FIGS. 2J and 2K show additional monomers in which R¹ and R² areindependently selected from the group consisting of, but not limited to,fluorine, alkyl, phenyl, alkyl-substituted phenyl, alkyl-substitutedfluorenyl, alkyl-substituted carbazolyl and alkyl-substitutedthiophenyl. Alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl, 7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substitutedthiophenyl can include 3-alkyl-substituted thiophenyl,4-alkyl-substituted thiophenyl, 5-alkyl-substituted thiophenyl,3,4-dialkyl-substituted thiophenyl, 3,5-dialkyl-substituted thiophenyland 4,5-dialkyl-substituted thiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher. R³ can be hydrogen, fluorine, trifluoro or —CN. FIG. 2L furthershows generic structures for both general monomers and narrow-bandmonomers present in a copolymer provided by the present invention. InFIG. 2L, each of R¹, R², R³ and R⁴ can be independently selected fromthe group consisting of, but not limited to, fluorine, alkyl, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl and alkyl-substituted thiophenyl. Alkyl substituted phenylcan include 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl,2,4-dialkylphenyl, 3,5-dialkylphenyl, and 3,4-dialkylphenyl.Alkyl-substituted fluorenyl can include 9,9-dialkyl-substitutedfluorenyl, 7-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substitutedthiophenyl can include 3-alkyl-substituted thiophenyl,4-alkyl-substituted thiophenyl, 5-alkyl-substituted thiophenyl,3,4-dialkyl-substituted thiophenyl, 3,5-dialkyl-substituted thiophenyland 4,5-dialkyl-substituted thiophenyl. The alkyl substituents includeC_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein nis 1 to 20. In some embodiments, n can be between 1 to 50 or higher.Each of R⁵ and R⁶ are independently selected from the group consistingof, but not limited to, fluorine, alkyl, phenyl, alkyl-substitutedphenyl, alkyl-substituted fluorenyl and alkyl-substituted carbazolyl.Alkyl substituted phenyl can include 2-alkylphenyl, 3-alkylphenyl,4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl, 7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. The alkylsubstituents include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher.

A variety of other BODIPY derivatives can be used for the presentinvention. BODIPY and BODIPY derivatives can be polymerized to formpolymers (e.g., homopolymers or heteropolymers) and/or can be attached(e.g., covalently attached) to a polymer backbone, sidechain and/orterminus. In some embodiments, the chromophoric polymer dots of thepresent invention can include a polymer that includes a narrow-bandmonomer having the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A) and R^(4B) isindependently selected from the group consisting of, but not limited to,hydrogen (H), deuterium (D), halogen, direct or branched alkyl,heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer, through at leastone attachment to R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), ora combination thereof. FIG. 4A shows examples of monomers that, e.g.,can be integrated with the polymer by attachment to R^(3A) and R^(3B)groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A) and R^(4B) isindependently selected from the group consisting of, but not limited to,hydrogen (H), deuterium (D), halogen, direct or branched alkyl,heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer through at least oneattachment to R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), or acombination thereof. The monomer can, for example, integrate with thebackbone of the polymer by attachment to the R^(3A) and R^(3B) groups.FIG. 4B shows examples of monomers that, e.g., can be integrated withthe polymer by attachment to R^(3A) and R^(3B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R¹, R^(2A) and R^(2B) is independently selected from thegroup consisting of, but not limited to, hydrogen (H), deuterium (D),halogen, direct or branched alkyl, heteroalkyl, heterocycloalkyl,heterocycloalkylene, alkoxy, aryl, hydroxyl, cyano, nitro, ether and itsderivatives, ester and its derivatives, alkyl ketone, alkylester,arylester, alkynyl, alkyl amine, fluoroalkyl, fluoroaryl, andpolyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy, and—(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted phenyl, pyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer through at least oneattachment, e.g., to R¹, R^(2A), R^(2B), or a combination thereof. Theparentheses indicate points of attachment of the monomer to the backboneof the polymer. FIG. 4C shows examples of monomers that, e.g., can beintegrated with the polymer (e.g., copolymerized in the polymer).

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), and R^(3B) is independentlyselected from the group consisting of, but not limited to, hydrogen (H),deuterium (D), halogen, direct or branched alkyl, heteroalkyl,heterocycloalkyl, heterocycloalkylene, alkoxy, aryl, hydroxyl, cyano,nitro, ether and its derivatives, ester and its derivatives, alkylketone, alkylester, arylester, alkynyl, alkyl amine, fluoroalkyl,fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy,and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-) substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl, pyrazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyrazolyl,oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedoxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted thiazolyl, imidazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted imidazolyl, pyrazinyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrazinyl,benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer through at least oneattachment to R¹, R^(2A), R^(2B), R^(3A), and R^(3B) or a combinationthereof. FIG. 4D shows examples of monomers that, e.g., can beintegrated with the polymer by attachment to R^(3A) and R^(3B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B),R^(5A), and R^(5B) is independently selected from the group consistingof, but not limited to, hydrogen (H), deuterium (D), halogen, direct orbranched alkyl, heteroalkyl, heterocycloalkyl, heterocycloalkylene,alkoxy, aryl, hydroxyl, cyano, nitro, ether and its derivatives, esterand its derivatives, alkyl ketone, alkylester, arylester, alkynyl, alkylamine, fluoroalkyl, fluoroaryl, and polyalkalene (e.g,mehtoxyethoxyethoxy, ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50),phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedphenyl, pyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted pyridyl, bipyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted bipyridyl tripyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted tripyridyl,furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedfuryl, thienyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted thienyl, pyrrolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyrrolyl, pyrazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrazolyl,oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedoxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted imidazolyl, pyrazinyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyrazinyl, benzooxadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzooxadizolyl, benzothiadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzothiadizolyl, fluorenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted fluorenyl,triphenylaminyl-substituted fluorenyl, diphenylaminyl-substitutedfluorenyl, alkyl-substituted carbazolyl, alkyl-substitutedtriphenylaminyl and alkyl-substituted thiophenyl. As exemplaryembodiments, alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl,3,4-dialkylphenyl; alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl, 6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., copolymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer through at least oneattachment to R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B),R^(5A), R^(5B), or a combination thereof. In certain embodiments, thenarrow-band monomers can be integrated into the backbone by attachmentto the R^(5A) and R^(5B) groups. FIG. 4E shows examples of monomersthat, e.g., can be integrated with the polymer by attachment to R^(5A)and R^(5B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R^(1A), R^(1B), R^(2A), R^(2B), R^(3A) and R^(3B) isindependently selected from the group consisting of, but not limited to,hydrogen (H), deuterium (D), halogen, direct or branched alkyl,heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer through at least oneattachment to R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B), or acombination thereof. FIG. 4F shows examples of monomers that, e.g., canbe integrated with the polymer by attachment to R^(1A), R^(1B), R^(2A),R^(2B), R^(3A) or R^(3B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), R^(5A)and R^(5B) is independently selected from the group consisting of, butnot limited to, hydrogen (H), deuterium (D), halogen, direct or branchedalkyl, heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer) through at leastone attachment to R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B),R^(5A), R^(5B), or a combination thereof. FIG. 4G shows examples ofmonomers that, e.g., can be integrated with the polymer by attachment toR^(5A) and R^(5B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein each of R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B), R^(4A)and R^(4B), is independently selected from the group consisting of, butnot limited to, hydrogen (H), deuterium (D), halogen, direct or branchedalkyl, heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl,and wherein each of R^(5A), R^(5B), R^(6A) and R^(6B) are independentlyselected from the group consisting of, but not limited to, hydrogen (H),deuterium (D), halogen, direct or branched alkyl, heteroalkyl,heterocycloalkyl, heterocycloalkylene, alkoxy, aryl, hydroxyl, cyano,nitro, ether and its derivatives, ester and its derivatives, alkylketone, alkylester, arylester, alkynyl, alkyl amine, fluoroalkyl,fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy,and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., copolymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer) through at leastone attachment to R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B),R^(4A), R^(4B), R^(5A), R^(5B), R^(6A), R^(6B) or a combination thereof.FIG. 4H shows examples of monomers that, e.g., can be integrated withthe polymer by attachment to R^(2A), R^(2B), R^(6A) or R^(6B) groups.

In some embodiments, the chromophoric polymer dots of the presentinvention can include a polymer that includes a narrow-band monomerhaving the formula:

wherein X represents aryl group and its derivatives, each of R¹, R², R³,R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is independentlyselected from the group consisting of, but not limited to, hydrogen (H),deuterium (D), halogen, direct or branched alkyl, heteroalkyl,heterocycloalkyl, heterocycloalkylene, alkoxy, aryl, hydroxyl, cyano,nitro, ether and its derivatives, ester and its derivatives, alkylketone, alkylester, arylester, alkynyl, alkyl amine, fluoroalkyl,fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy,and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.When X represents naphthalene and its derivatives, the narrow-bandmonomer can be integrated into a backbone (e.g., polymerized in thepolymer) and/or covalently attached to the backbone, a terminus, or asidechain of the polymer) of the polymer through at least one attachmentto R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹² or a combination thereof. When X representsanthracene and its derivatives, the narrow-band monomer can beintegrated into a backbone of the polymer and/or covalently attached tothe backbone, a terminus, or a sidechain of the polymer through at leastone attachment to R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵ or acombination thereof.

Narrow band monomers of the present invention can further includedipyrrin derivatives. Dipyrrin and dipyrrin derivatives can bepolymerized to form polymers (e.g., homopolymers or heteropolymers)and/or can be attached (e.g., covalently attached) to a polymerbackbone, sidechain and/or terminus. For example, the chromophoricpolymer dots of the present invention can include a polymer thatincludes a narrow-band monomer having the formula:

wherein M is a metal. Examples of M can be, but not limited to, Na, Li,Zn, Co, or Si. X can include substituents such as, but not limited to,halogen, alkyl, phenyl, alkylphenyl, thiophenyl, alkylthiophenyl,alkoxyl, alkoxylphenyl, alkylthiophenyl, ester, or hydroxyl. The numberof X groups (n) can be 1 or more than 1, and n can be 0, 1, 2, 3, 4.Each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), R^(5A), andR^(5B) can be independently selected from the group consisting of, butnot limited to, hydrogen (H), deuterium (D), halogen, direct or branchedalkyl, heteroalkyl, heterocycloalkyl, heterocycloalkylene, alkoxy, aryl,hydroxyl, cyano, nitro, ether and its derivatives, ester and itsderivatives, alkyl ketone, alkylester, arylester, alkynyl, alkyl amine,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-) substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl,N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, and N-dialkoxyphenyl-4-phenyl.The narrow-band monomer can be integrated into a backbone of the polymer(e.g., polymerized in the polymer) and/or covalently attached to thebackbone, a terminus, or a sidechain of the polymer) through at leastone attachment to R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B),R^(5A), R^(5B), or a combination thereof.

Additional examples of dipyrrin derivatives are shown in FIG. 5, whereinX can include substituents such as, but not limited to, halogen, alkyl,phenyl, alkylphenyl, thiophenyl, alkylthiophenyl, alkoxyl, ester, orhydroxyl. The number of X groups (n) can be 1 or more than 1, and n canbe 0, 1, 2, 3, 4.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude squaraine and squaraine derivatives as narrow-band monomers.Squaraine derivatives include but are not limited to their alkylderivatives, aryl derivatives, alkyne derivatives, aromatic derivatives,alkoxide derivatives, aza derivatives, their extended systems andanalogues. The narrow-band emissive polymers can also include any othermonomers. The squaraine and their derivatives can be energy acceptorsand other monomers can be energy donors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

Suitable squaraine derivatives for use in the present invention caninclude the following structures described below. Squaraine andsquaraine derivatives can be polymerized to form polymers (e.g.,homopolymers or heteropolymers) and/or can be attached (e.g., covalentlyattached) to a polymer backbone, sidechain and/or terminus. Chromophoricpolymer dots of the present invention can include a polymer thatincludes a narrow-band monomer having the formula:

wherein each of X¹ and X² is independently selected from the groupconsisting of oxygen, sulfur and nitrogen; each of R^(1A) and R^(1B) isindependently selected from the group consisting of, but not limited,alkylene, alkenylene, arylene, heteroarylene, phenylene, azulene,cycloalkylene, and heterocycloalkylene; and each of R^(2A) and R^(2B) isa reactive group independently selected from the group consisting of,but not limited to, a halide, hydroxyl, and amino. Other reactive groupscan be used. In some embodiments, the halide is a chloro, a bromo, or aniodo group. The reactive group can be used to integrate the monomer intoa polymer, e.g. along the backbone of the polymer (e.g., by polymerizingin the polymer) and/or to attach the monomer by covalent attachment tothe backbone, a terminus, or a sidechain of the polymer.

The present invention can include oxygen-containing squarainederivatives. Chromophoric polymer dots of the present invention caninclude a polymer that includes a narrow-band monomer having theformula:

wherein each of R^(1A) and R^(1B) is independently selected from thegroup consisting of, but not limited to, alkylene, alkenylene, arylene,heteroarylene, phenylene, azulene, cycloalkylene, andheterocycloalkylene; and each of R^(2A) and R^(2B) is a reactive groupindependently selected from the group consisting of, but not limited to,a halide, hydroxyl, and amino. Other reactive groups can be used. Insome embodiments, the halide is a chloro, a bromo, or an iodo group. Thereactive group can be used to integrate the monomer into a polymer (e.g.along the backbone of the polymer by polymerizing in the polymer) and/orto attach the monomer by covalent attachment to the backbone, aterminus, or a sidechain of the polymer.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A) and R^(1B) is independently selected from thegroup consisting of, but not limited to, hydrogen, methyl, alkyl,phenyl, araalkyl, and alkoxy-phenyl; each of R^(2A) and R^(2B) isindependently selected from the group consisting of, but not limited to,hydrogen, methyl, alkyl, phenyl, araalkyl, and alkoxy-phenyl; each ofR^(3A) and R^(3B) is a reactive group independently selected from thegroup consisting of, but not limited to, chloro, bromo, iodo, andhydroxyl; each of R^(4A) and R^(4B) is independently is selected from agroup consisting of, but not limited to, hydroxyl, hydrogen, alkyl,phenyl, araalkyl, and alkoxy-phenyl; and each of R^(5A) and R^(5B) isindependently selected from the group consisting of, but not limited to,hydrogen, methyl, alkyl, phenyl, araalkyl, and alkoxy-phenyl. Otherreactive groups can be used. The reactive group can be used to integratethe monomer into a polymer (e.g. along the backbone of the polymer bypolymerizing in the polymer) and/or to attach the monomer by covalentattachment to the backbone, a terminus, or a sidechain of the polymer.FIG. 6A shows some examples of narrow band monomers that can beintegrated into the polymer by reaction with the reactive groups, e.g.,I or Br.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A) and R^(1B) is a reactive group independentlyselected from the group consisting of, but not limited to, chloro,bromo, iodo, and hydroxyl; and each of R^(2A) and R^(2B) is selectedfrom the group consisting of, but not limited to, hydrogen, methyl,alkyl, phenyl, araalkyl, and alkoxy-phenyl. Other reactive groups can beused. The reactive group can be used to integrate the monomer into apolymer (e.g. along the backbone of the polymer by polymerizing in thepolymer) and/or attach the monomer by covalent attachment to thebackbone, a terminus, or a sidechain of the polymer. FIG. 6B shows anexample monomer that can be integrated into the polymer by reaction withthe reactive groups, e.g., Br.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of X¹ and X² is independently selected from the groupconsisting of carbon, sulphur, and selenium; each of R^(1A) and R^(1B)is a reactive group independently selected from the group consisting of,but not limited to, chloro, bromo, iodo, and hydroxyl; and each ofR^(2A) and R^(2B) is independently selected from the group consistingof, but not limited to, hydrogen, methyl, alkyl, phenyl, araalkyl,alkoxy-phenyl, N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, andN-dialkoxylphenyl-4-phenyl. Other reactive groups can be used. Thereactive group can be used to integrate the monomer into a polymer (e.g.along the backbone of the polymer by polymerizing in the polymer) and/orattach the monomer by covalent attachment to the backbone, a terminus,or a sidechain of the polymer. FIG. 6B shows example monomers that canbe integrated into the polymer by reaction with the reactive groups,e.g., Br.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(2A) and R^(2B) is a reactive group independentlyselected from the group consisting of, but not limited to, chloro,bromo, iodo, and hydroxyl; and each of R^(1A) and R^(1B) is selectedfrom the group consisting of, but not limited to, hydrogen, methyl,alkyl, phenyl, araalkyl, alkoxy-phenyl, N-dialkyl-4-phenyl,N-diphenyl-4-phenyl, and N-dialkoxylphenyl-4-phenyl. Other reactivegroups can be used. The reactive group can be used to integrate themonomer into a polymer (e.g. along the backbone of the polymer bypolymerizing in the polymer) and/or attach the monomer by covalentattachment to the backbone, a terminus, or a sidechain of the polymer.

The present invention can include sulphur-containing squarainederivatives. Chromophoric polymer dots of the present invention caninclude a polymer that includes a narrow-band monomer having theformula:

wherein each of R^(1A) and R^(1B) is independently selected from thegroup consisting of, but not limited to, alkylene, alkenylene, arylene,heteroarylene, phenylene, azulene, cycloalkylene, andheterocycloalkylene; and each of R^(2A) and R^(2B) is a reactive groupindependently selected from the group consisting of, but not limited to,a halide, hydroxyl, and amino. In some embodiments, the halide is achloro, a bromo, or an iodo group. Other reactive groups can be used.The reactive group can be used to integrate the monomer into a polymer(e.g. along the backbone of the polymer by polymerizing in the polymer)and/or attach the monomer by covalent attachment to the backbone, aterminus, or a sidechain of the polymer.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of X¹ and X² is independently selected from the groupconsisting of carbon, sulphur, and selenium; each of R^(1A) and R^(1B)is a reactive group independently selected from the group consisting of,but not limited to, chloro, bromo, iodo, and hydroxyl; and each ofR^(2A) and R^(2B) is independently selected from the group consistingof, but not limited to, hydrogen, methyl, alkyl, phenyl, araalkyl,alkoxy-phenyl, N-dialkyl-4-phenyl, N-diphenyl-4-phenyl, andN-dialkoxylphenyl-4-phenyl. Other reactive groups can be used. FIG. 6Cshows example monomers that can be integrated into the polymer byreaction with the reactive groups, e.g., Br (e.g. along the backbone ofthe polymer by polymerizing in the polymer) and/or attached by covalentattachment to the backbone, a terminus, or a sidechain of the polymer).

The present invention can include nitrogen-containing squarainederivatives. Chromophoric polymer dots of the present invention caninclude a polymer that includes a narrow-band monomer having theformula:

wherein each of R^(1A) and R^(1B) is independently selected from thegroup consisting of, but not limited to, alkylene, alkenylene, arylene,heteroarylene, phenylene, azulene, cycloalkylene, andheterocycloalkylene; each of R^(2A) and R^(2B) is a reactive groupindependently selected from the group consisting of, but not limited to,a halide, hydroxyl, and amino; and each of R^(3A) and R^(3B) isindependently selected from the group consisting of hydrogen, methyl,alkyl, phenyl, aralkyl, and alkoxy-phenyl. Other reactive groups can beused. In some embodiments, the halide is a chloro, a bromo, or an iodogroup. The reactive group can be used to integrate the monomer alonginto a polymer (e.g. along the backbone of the polymer by polymerizingin the polymer) and/or attach the monomer by covalent attachment to thebackbone, a terminus, or a sidechain of the polymer. FIG. 6D showsexample monomers that can be integrated into the polymer by reactionwith the reactive groups, e.g., Br.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A), R^(1B), R^(2A) and R^(2B) are independentlyselected from the group consisting of, but not limited to, hydrogen,deuterium, alkyl, aryl, acetyl, and hydroxyl; and each of R^(3A),R^(3B), R^(4A) and R^(4B) are independently selected from the groupconsisting of, but not limited to, hydrogen, deuterium, alkyl, aryl,amino, sulfide, aldehyde, ester, ether, acid, hydroxyl, and halide. Thenarrow-band monomer can be integrated into a backbone of the polymer(e.g. along the backbone of the polymer by polymerizing in the polymer)and/or attached by covalent attachment to the backbone, a terminus, or asidechain of the polymer through at least one attachment to R^(1A),R^(1B), R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B) or a combinationthereof.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A), R^(1B), R^(2A) and R^(2B) are independentlyselected from the group consisting of, but not limited to, hydrogen,deuterium, alkyl, aryl, acetyl, and hydroxyl; and each of R^(3A),R^(3B), R^(4A), R^(4B), R^(5A), R^(5B), R^(6A) and R^(6B) areindependently selected from the group consisting of, but not limited to,hydrogen, deuterium, alkyl, aryl, amino, sulfide, aldehyde, ester,ether, acid, hydroxyl, and halide. The narrow-band monomer can beintegrated into a backbone of the polymer (e.g. along the backbone ofthe polymer by polymerizing in the polymer) and/or attached by covalentattachment to the backbone, a terminus, or a sidechain of the polymerthrough at least one attachment to R^(1A), R^(1B), R^(2A), R^(2B),R^(3A), R^(3B), R^(4A), R^(4B), R^(5A), R^(5B), R^(6A), R^(6B) or acombination thereof.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A), R^(1B), R^(1C), R^(2A), R^(2B), R^(2C), R^(3A),R^(3B), R^(3C), R^(4A), R^(4B), R^(4C), R^(5A), R^(5B), R^(5C), R^(6A),R^(6B), and R^(6C) are independently selected from the group consistingof, but not limited to, hydrogen, deuterium, alkyl, aryl, amino,sulfide, aldehyde, ester, ether, acid, hydroxyl, and halide, and each ofR^(7A), R^(7B), and R^(7C) is independently selected from the groupconsisting of, but not limited to, hydrogen, deuterium, alkyl, aryl andacetyl. The narrow-band monomer can be integrated into a backbone of thepolymer (e.g. along the backbone of the polymer by polymerizing in thepolymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentto R^(1A), R^(1B), R^(1C), R^(2A), R^(2B), R^(2C), R^(3A), R^(3B),R^(3C), R^(4A), R^(4B), R^(4C), R^(5A), R^(5B), R^(5C), R^(6A), R^(6B),R^(6C), R^(7A), R^(7B) or a combination thereof. Alternatively, as shownhere, the monomer described herein can be integrated with the polymer byattachment as shown by the parentheses.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R^(1A) and R^(1B) is independently selected from thegroup consisting of, but not limited to hydrogen, deuterium, alkyl andaryl; and each of R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), R⁵,R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹ and R¹² are independently selected from thegroup consisting of, but not limited to, hydrogen, deuterium, alkyl,aryl, cyano, amino, sulfide, aldehyde, ester, ether, acid, hydroxyl, andhalide. The narrow-band monomer can be integrated into a backbone of thepolymer (e.g. along the backbone of the polymer by polymerizing in thepolymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentto R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B), R^(4A), R^(4B), R⁵,R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹² or a combination thereof.

Chromophoric polymer dots of the present invention can include a polymerthat includes a narrow-band monomer having the formula:

wherein each of R¹³ is independently selected from the group consistingof, but not limited to, hydrogen, deuterium, alkyl and aryl; and R¹⁴,R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³ and R²⁴ are independentlyselected from the group consisting of, but not limited to, hydrogen,deuterium, alkyl, aryl, cyano, amino, sulfide, aldehyde, ester, ether,acid, hydroxyl, and halide. The narrow-band monomer can be integratedinto a backbone of the polymer (e.g. along the backbone of the polymerby polymerizing in the polymer) and/or attached by covalent attachmentto the backbone, a terminus, or a sidechain of the polymer through atleast one attachment to R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³ and R²⁴ or a combination thereof. FIG. 6E shows examplemonomers that can be integrated into the polymer by reaction with thereactive groups, e.g., Br.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude metal complexes and their derivatives as narrow-band monomers.Metal complexes and their derivatives include but are not limited totheir alkyl derivatives, aryl derivatives, alkyne derivatives, aromaticderivatives, alkoxide derivatives, aza derivatives, their extendedsystems and analogues. The narrow-band emissive polymers can alsoinclude any other monomers. The metals can be any metal such as Na, Li,Zn, Mg, Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt, Pd, Ru, Rh, Re, Os, Ir,Ag, Au and so on. The metal complexes can be energy acceptors and othermonomers can be energy donors so that the final Pdots can exhibitnarrow-band emissions. The narrow-band emissive chromophoric polymers ingood solvents may exhibit broad emissions or narrow emissions. However,their nanoparticle form gives narrow-band emissions. The emission FWHMof the above Pdots is less than 70 nm. In certain embodiments, the FWHMis less than 60 nm, less than 50 nm, less than 40 nm, less than 30 nm,or less than 20 nm.

Examples of metal complexes and metal complex derivatives are shown inFIGS. 7A-7C. Metal complexes and metal complex derivatives can bepolymerized to form polymers (e.g., homopolymers or heteropolymers)and/or can be attached (e.g., covalently attached) to a polymerbackbone, sidechain and/or terminus. As shown in FIG. 7A, the metalcomplexes of the present invention include derivatives of the metalcomplexes. The metal complex monomers shown in FIG. 7A can include thecompounds as shown, wherein R¹ and R² are independently selected fromthe group consisting of, but not limited to, phenyl, alkyl-substitutedphenyl, alkyl-substituted fluorenyl, diphenyl-substituted fluorenyl,triphenylaminyl-substituted fluorenyl, diphenylaminyl-substitutedfluorenyl, alkyl-substituted carbazolyl, alkyl-substitutedtriphenylaminyl and alkyl-substituted thiophenyl. Alkyl substitutedphenyl can include 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl,2,4-dialkylphenyl, 3,5-dialkylphenyl, and 3,4-dialkylphenyl.Alkyl-substituted fluorenyl can include 9,9-dialkyl-substitutedfluorenyl, 7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-dialkyl-substitutedtriphenylaminyl. Alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. As will be further understood by oneof ordinary skill in the art, the general monomer (D) and the narrowband metal complex monomers are present in the polymer at a ratio whereD is present as x and the narrow band monomer is present as 1−x. Forexample, D may be present at 90% or x=0.9 and the narrow band monomer ispresent at 10% or 1−x=0.1. FIGS. 7B and 7C show additional examplemonomers for use in the present invention.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude porphyrin, metalloporphyrin, and their derivatives asnarrow-band monomers. Porphyrin, metalloporphyrin, and their derivativescan be polymerized to form polymers (e.g., homopolymers orheteropolymers) and/or can be attached (e.g., covalently attached) to apolymer backbone, sidechain and/or terminus. Porphyrin,metalloporphyrin, and their derivatives include but are not limited totheir alkyl derivatives, aryl derivatives, alkyne derivatives, aromaticderivatives, alkoxide derivatives, aza derivatives, their extendedsystems and analogues. The metals in the metalloporphyrins can be anymetal such as Na, Li, Zn, Mg, Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt,Pd, Ru, Rh, Re, Os, Ir, Ag, Au and so on. The narrow-band emissivepolymers can also include any other monomers. The porphyrin,metalloporphyrin and their derivatives can be energy acceptors and othermonomers can be energy donors so that the final Pdots can exhibitnarrow-band emissions. The narrow-band emissive chromophoric polymers ingood solvents may exhibit broad emissions or narrow emissions. However,their nanoparticle form gives narrow-band emissions. The emission FWHMof the above Pdots is less than 70 nm. In certain embodiments, the FWHMis less than 60 nm, less than 50 nm, less than 40 nm, less than 30 nm,or less than 20 nm.

FIG. 8 shows example porphyrin and porphyrin derivatives for use in thepresent invention. As shown in FIG. 8, the porphyrin derivatives cancomplex, e.g., with Pt and Zn. Also, R¹ and R² can be independentlyselected from the group consisting of, but not limited to, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl, alkyl-substitutedthiophenyl, fluorine (F), cyano (CN) and trifluoro (CF₃). Alkylsubstituted phenyl can include 2-alkylphenyl, 3-alkylphenyl,4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl and 6-alkyl-9,9-dialkyl-substituted fluorenyl.Alkyl-substituted carbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, and 4-alkylthiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃,wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher. The narrow-band monomer can be integrated into a backbone of thepolymer (e.g., by copolymerizing in the polymer) and/or attached bycovalent attachment to the backbone, a terminus, or a sidechain of thepolymer, through at least one attachment to R¹, R², or a combinationthereof. Alternatively, as shown in FIG. 8, the monomers describedherein can be integrated with the polymer by attachment as shown bybrackets. As described herein, the general monomer (D) and the narrowband metal complex monomers are present in the polymer at a ratio whereD is present as x and the narrow band monomer is present as 1−x.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude phthalocyanine and its derivatives as monomers. Phthalocyanineand its derivatives as monomers can be polymerized to form polymers(e.g., homopolymers or heteropolymers) and/or can be attached (e.g.,covalently attached) to a polymer backbone, sidechain and/or terminus.Phthalocyanine derivatives include but are not limited to their alkylderivatives, aryl derivatives, alkyne derivatives, aromatic derivatives,alkoxide derivatives, aza derivatives, their extended systems andanalogues. The metals in the phthalocyanine derivatives can be any metalsuch as Na, Li, Zn, Mg, Fe, Mn, Co, Ni, Cu, In, Si, Ga, Al, Pt, Ru, Rh,Re, Os, Ir, Ag, Au or Pd. The narrow-band emissive polymers can alsoinclude any other monomers. The phthalocyanine derivatives can be energyacceptors so that the final Pdots can exhibit narrow-band emissions. Thenarrow-band emissive chromophoric polymers in good solvents may exhibitbroad emissions or narrow emissions. However, their nanoparticle formgives narrow-band emissions. The emission FWHM of the above Pdots isless than 70 nm. In certain embodiments, the FWHM is less than 60 nm,less than 50 nm, less than 40 nm, less than 30 nm, or less than 20 nm.

FIG. 9 shows an example phthalocyanine structure that can be used as anarrow band monomer in the present invention. As shown, M can be, but isnot limited to, Cu, Zn, Mn, Fe, Si, Pt, Co, Ca, Ni, Na, Mg Ru, Rh, Re,Os, Ir, Ag, Au, Pd or Al. R¹, R², R³, and R⁴ can be a substituentattached to any suitable position in the isoindole portions of themonomer. The narrow-band monomer can be integrated into a backbone ofthe polymer (e.g., by copolymerizing in the polymer) and/or attached bycovalent attachment to the backbone, a terminus, or a sidechain of thepolymer through at least one attachment to R¹, R², R³, R⁴, or acombination thereof. In certain embodiments, each of R¹, R², R³, and R⁴can be independently selected from the group consisting of, but notlimited to, phenyl, alkyl-substituted phenyl, alkyl-substitutedfluorenyl, alkyl-substituted carbazolyl, alkyl-substitutedtriphenylaminyl, alkyl-substituted thiophenyl, hydrogen, fluorine (F),cyano (CN) and trifluoro (CF₃). Alkyl substituted phenyl can include2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl. Alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl and6-alkyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substituted carbazolylcan include N-alkyl-substituted carbazolyl, 6-alkyl-substitutedcarbazolyl and 7-alkyl-substituted carbazolyl. Alkyl-substitutedthiophenyl can include 2-alkylthiophenyl, 3-alkylthiophenyl, and4-alkylthiophenyl. The alkyl substituents can include C_(n)H_(2n+1), orC_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. Insome embodiments, n can be between 1 to 50 or higher.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude Lanthanide complexes and their derivatives as narrow-bandmonomers. Lanthanide complexes and lanthanide complex derivatives asmonomers can be polymerized to form polymers (e.g., homopolymers orheteropolymers) and/or can be attached (e.g., covalently attached) to apolymer backbone, sidechain and/or terminus. Lanthanide complexes andtheir derivatives include but are not limited to their alkylderivatives, aryl derivatives, alkyne derivatives, aromatic derivatives,alkoxide derivatives, aza derivatives, their extended systems andanalogues. The narrow-band emissive polymers can also include any othermonomers. The Lanthanide complexes and their derivatives can be energyacceptors and other monomers can be donors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, less than 20 nm, or less than 10 nm.

The lanthanide complexes described herein can have different narrowemission properties and mechanisms as compared to transition metalcomplexes. For example, a fluorescence mechanism of lanthanide (III)complexes (such as Ce (III), Pr(III), Nd(III), Sm(III), Eu(III),Tb(III), Dy(III), Ho(III), Er(III), Tm(III), or Yb(III)), whose 4fexcited states are not empty and not fully filled by the electron, canbe that energy absorbed by the organic ligand is first transferred fromthe singlet state to the triplet state (intersystem crossing) of theligand, and then transferred (or is directly transferred from thesinglet state of the organic ligand) to the 4f excited states of thelanthanide ions by the resonant energy transfer process. Here, theemission comes from the 4f-4f transitions of the Ln(III) ions. Due tothe inner shell f orbital electrons being shielded from the environmentby the filled 5S²5p⁶ sub-shells, they don't vary much with theenvironment. Consequently, the inner-shell 4f-4f transitions are sharpand result in narrow emission. For some divalent lanthanide ions, suchas Sm(II), Eu (II), and YB (II), the emission is from the 5d-4ftransitions. These properties of Lanthanide containing Pdotssurprisingly can provide additional features as compared to othernon-Lanthanide metal based Pdots, such as Pdots containing transitionmetal complexes.

In some embodiments, lanthanide complexes that can be used with thepresent invention can be described with the following formula:

For example, the lanthanide complexes can be used as the narrow-bandemissive monomers. The lanthanide complexes can, e.g., be repeatingunits of the polymer backbone. The lanthanide complexes can, e.g., beattached to the side-chains of the polymer. The lanthanide complexescan, e.g., be attached to the terminus of the polymer. Ln is lanthanidemetal ion which has an unfilled inner shell and can accept energy fromthe organic ligands or general polymers to give narrow-band emission.There are several lanthanides that can be used, which can be the same ordifferent, and can be selected from, e.g., Ce(III), Pr(III), Nd(III),Sm(III), Sm(II) Eu(III), Eu(II), Tb(III), Dy(III), Ho(III), Er(III),Tm(III), Yb(III), Yb(II). As shown in the formula above, L₁ and L₂ canbe organic ligands. The ligands can be the same or different. There canbe a plurality of ligands L₁. L₁ that can be anionic ligands coordinatedwith Ln. The total valences of L₁ and Ln can be equal to finally form aneutral lanthanide complex. In addition to acting as anionic ligands, L₁can include additional groups to act as neutral ligands to becoordinated with Ln. L₁ can be monodentate, bidentate or polydentate andthere can be one or more ligands L₁ in the lanthanide complex. In someembodiments, L₁ can be a bridged ligand and be coordinated with Ln toform binuclear, trinuclear and polynuclear lanthanide complexes. Some ofbridged L₁ can form cryptands and can be coordinated to Ln to synthesizelanthanide cryptate. L₂ can be a neutral ligand. There can be aplurality of ligands L₂. L₂ can be the same or different. L₂ can bemonodentate, bidentate or polydentate and there can be one or moreligands L₂ in the lanthanide complex. In some embodiments, L₂ can be abridged ligand and be coordinated with Ln to form binuclear, trinuclearand polynuclear lanthanide complexes. Some of bridged L₂ can formcryptands and can be coordinated to Ln to synthesize lanthanidecryptate.

FIGS. 10A-10H shows example derivatives of lanthanide complexes that canbe used as narrow band emissive units for the present invention. In someembodiments, the general monomers described herein can be designed toact as energy donors that transfer energy to selected lanthanidecomplexes. For example, the emission profile of a general monomer canoverlap with the absorption profile of a lanthanide complex. Throughenergy transfer, the lanthanide complex can be excited and then emitlight with a narrow band emission (e.g., of less than 70 nm FWHM). Theenergy transfer can be accomplished through a variety of ways. In someembodiments, homopolymers (e.g. P1-P10) or heteropolymers includinggeneral monomers (e.g., D1, D2, and/or monomers of homopolymer inP1-P10, and/or M1-M10) can be chemically bonded with lanthanidecomplexes and condensed into polymer dots. In certain embodiments,homopolymers (e.g. P1-P10) or heteropolymers including general monomers(e.g., D1, D2, and/or monomers of homopolymer in P1-P10, and/or M1-M10)can be chemically crosslinked with homo- or hetero-polymers includinglanthanide complexes and condensed into polymer dots. In someembodiments, the general monomers (e.g., D1, D2, and/or monomers ofhomopolymer in P1-P10, and/or M1-M10) can be copolymerized in anycombination with the lanthanide complexes. Due at least in part topacking of the polymers and the lanthanide complexes, energy transfercan be achieved between the general monomers and the lanthanidecomplexes.

The lanthanide complexes can be added to polymers in a variety of ways.For example, FIG. 10A shows schematic structures of narrow-band emissivepolymers that can include lanthanide complexes. Lanthanide complexes canbe used to produce a narrow-band emissive homopolymer. Lanthanidecomplexes can also be copolymerized with any of the general polymers tosynthesize a narrow-band emissive copolymer or multi-componentheteropolymer. Lanthanide complexes can be used as a narrow-bandemissive unit to cross-link with the side-chains of conventionalsemiconducting polymers to form narrow-band emissive polymers. In someembodiments, the present invention can include lanthanide complexes asmonomers in a homopolymer or heteropolymer. In certain embodiments, thelanthanide complexes can be linked (e.g., covalently bonded) to aside-chain of a homopolymer or heteropolymer having general monomersthat, e.g., can act as energy donors to the lanthanide complexes (e.g.,D1 and/or D2 and/or D2′ monomers and/or monomers of homopolymer inP1-P10, and/or M1-M10 described herein can be the general monomers). Thelanthanide complexes can also be copolymerized with a general monomer toform a heteropolymer. In each embodiment, the general monomer can beluminescent or non-luminescent. In some embodiments, the general monomermay absorb energy and transfer the energy, either directly or indirectly(e.g. via cascade energy transfer as described elsewhere in thisapplication), to the lanthanide complexes. The length of the polymers asdenoted, e.g., by n, in FIG. 10A can be designed to have any suitablelength to result in any suitable polymer molecular weight (Mw). Mw, forexample, can range between 500 and 1,000,000.

FIG. 10B shows example derivatives of lanthanide complexes that can beused as narrow band monomers for the present invention. A variety ofelements in the Lanthanide series can be used, such as Europium (Eu) andTerbium (Tb). The lanthanide complex monomers shown in FIG. 10B caninclude the monomers as shown, wherein R¹ and R² are independentlyselected from the group consisting of, but not limited to, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl,diphenyl-substituted fluorenyl, triphenylaminyl-substituted fluorenyl,diphenylaminyl-substituted fluorenyl, alkyl-substituted carbazolyl,alkyl-substituted triphenylaminyl and alkyl-substituted thiophenyl.Alkyl substituted phenyl can include 2-alkylphenyl, 3-alkylphenyl,4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, 3,4-dialkylphenyl.Alkyl-substituted fluorenyl can include 9,9-dialkyl-substitutedfluorenyl, 7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl. Alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl. Alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. As will be further understood by oneof ordinary skill in the art, the general monomer (D) and the narrowband metal complex monomers are present in the polymer at a ratio whereD is present as x and the narrow band monomer is present as 1−x. Forexample, D may be present at 90% or x=0.9 and the narrow band monomer ispresent at 10% or 1−x=0.1.

In some embodiments, the lanthanide complexes can be used as anarrow-band emissive unit to chemically crosslink with conventionalsemiconducting polymers to form narrow-band emissive polymer dots. Thegeneral semiconducting polymers can absorb energy and transfer theenergy to the lanthanide complexes. FIG. 10C shows non-limiting examplesof general homopolymers (e.g., P1-P10) and examples of generalheterpolymers including one type of monomers of homopolymer P1-P10 andother types of monomers (e.g., M1-M10). The general polymers can bedesigned to act as energy donors to selected lanthanide complexes. Insome embodiments, homo- or heteropolymers including general monomers(e.g., monomers of homopolymers P1-P10, and/or monomers of M1-M10) canbe chemically bonded with lanthanide complexes and condensed intopolymer dots. In certain embodiments, homo- or heteropolymers includinggeneral monomers (e.g., monomers of homopolymers P1-P10, and/or monomersof M1-M10) can be physically blended or chemically crosslinked withhomo- or hetero-polymers with attached lanthanide complexes andcondensed into polymer dots. In some embodiments, the general monomers(e.g monomers of homopolymers P1-P10, and/or monomers of M1-M10) can becopolymerized in any combination with the lanthanide complexes. Due atleast in part to packing of the polymers and the lanthanide complexes,energy transfer can be achieved between the general polymers and thelanthanide complexes.

Some example general polymers that can transfer energy to lanthanidecomplexes are shown in FIG. 10C. For example, P1-P10 are homopolymers ofthe various monomers having a defined length, n. With respect tohomopolymers of P1-P10 in FIG. 10C, each of R¹, R², R³ and R⁴ can beindependently selected from, but not limited to, the group consisting ofH, D, F, Cl, Br, I, alkoxy, aryloxy, alkyl, aryl, alkylketone,arylketone, alkylester, arylester, amide, carboxylic acid, fluoroalkyl,fluoroaryl, and polyalkaleneoxy. In some embodiments, two of the Rgroups (e.g., R¹, R², R³ and R⁴) may be bridging (e.g., covalentlylinked together to form a cyclic group, such as a cycloalkyl,hetercycloalkyl, aryl, or heteraryl group). Each of X and Z can beindependently selected from the group consisting of —O—, —S—, —N—,—NR⁵—, —PR⁵— and —CR⁵R⁶—, —CR⁵R⁶CR⁷CR⁸—, —N═CR⁵—, —CR⁵═CR⁶—, —N═N—, and—(CO)—, wherein each of R⁵, R⁶, R⁷ and R⁸ can be independently selectedfrom, but not limited to, the group consisting of H, D, F, Cl, Br, I,alkoxy, aryloxy, alkyl, aryl, alkyleneoxy, polyalkaleneoxy, alkoxy,aryloxy, fluoroalkyl and fluoroaryl. Any two of R⁵, R⁶, R⁷ and R⁸ may bebridging. In certain embodiments, the polymers include general monomersthat can be aromatic and have appropriate conjugation lengths leading tohigh extinction coefficient in typical wavelength in the region ofultraviolet (UV) to near infrared (NIR), i.e., 200-1800 nm, leading totypical fluorescence in the region of 200-1800 nm. The polymers can begood donors for transferring energy, either directly or indirectly (e.g.by cascade energy transfer), to lanthanide complexes. The generalmonomers can be included in polymers that are homopolymers, orcopolymers, or heteropolymers that contain more than two types ofmonomers. These polymers may be linear, branched, hyperbranched,dendritic, crosslinked, random, block, graft, or any structural type. Asalso shown in FIG. 10C, some example general polymers include othertypes of monomers (M1-M10). The copolymer of P (a general monomer) and M(another general monomer) can be mixed in a suitable ratio, and ascharacterized by x and 1−x. As also shown, M8 and M9 can be substitutedwith a range of alkyl groups defined by the formula C_(n)H_(2n+1) and0<n<20.

A variety of ligands can be used for the lanthanide complexes.Additional examples are shown, e.g., in FIG. 10D. FIG. 10D showsnon-limiting examples of anionic ligands L₁ that can include β-diketone,pyrazolone, isoxazolone, carboxylic acid, phthalocyanine,8-hydroxyquinoline, pyrazol borate, porphyrin, salicylaldehyde,phenylsalicylaldehyde, adenine, purine,2-(2-hydroxyphenyl)benzothiadiazole, 2-(2-hydroxyphenyl)quinolone,1-naphthol-2-carboxaldehyde, hydroxybenzophenone, 1,2-dihydroxybenzene,dihydroxynaphthalene, droxylfluorenone, 7-hydroxyinden-1-one,7-hydroxy-3-phenylinden-1-one, 2-hydroxy-dimethylbenzene-1,3-diamide,1,8-bis(4-methyl-2-hydroxy-benzamido)-3,6-dioxaoctane,2-hydroxy-N-methylbenzamide, bis(2-hydroxy-N-methylbenzamide), andtri(2-hydroxy-N-methylbenzamide), 8-hydroxyquinazoline,8-hydroxyquinoxaoline, hydroxybenzoxazole, hydroxy-2-phenyl benzoxazole,hypoxanthine, and the like. In some embodiments, L₁ has at least onearomatic ring. In some embodiments, L₁ has a direct chemical bondbetween two of the aryls groups to form a biaryl group, or has two ringsin a fused ring system. The ligand can also have a multi-aryl group or afused multi-ring group. The functional aryl groups can be bridged byalkyl, aryl, amine and other groups to form (semi-)macrocyclic ligands.In some embodiments, some of the bridged L₁ ligands can form cryptandsand can be coordinated to Ln to synthesize lanthanide cryptate. In someembodiments, the ligand can include aromatic ring member groups (theanionic-neutral ligand II in FIG. 10F). As shown, A₁-A₄ can be ringmembers and independently selected from substituted or unsubstitutedaryl and heteroaryl (e.g., azulene) moieties. In some embodiments, eachR′, R″ and R′″ groups can be independently selected from H, substitutedor unsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted heteroaryl, halogen, cyano (CN),substituted or unsubstituted aryl, substituted or unsubstitutedfluoroalkyl, substituted or unsubstituted fluoroaryl, substituted orunsubstituted alkoxy, substituted or unsubstituted heteroaryl and acyl.

FIG. 10E shows non-limiting examples of some substituted groups in theanionic ligands L₁. In some embodiments, R₁-R_(40+n) can beindependently selected from, but not limited to, H, D, halogen, director branched alkyl, alkoxy, aryl, alkyl ketone, alkylester, arylester,amide, fluoroalkyl, fluoroaryl, and polyalkalene (e.g,mehtoxyethoxyethoxy, ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50),phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedphenyl, pyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted bipyridyl tripyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl,pyrrolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrrolyl, pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher.

FIG. 10F shows non-limiting examples of neutral ligands L₂. In someembodiments, L₂ can include substituted or unsubstituted pyridine,substituted or unsubstituted bipyridine, substituted or unsubstitutedtripyridine, substituted or unsubstituted 1,10-phenanthroline,substituted or unsubstituted phosphine oxide, substituted orunsubstituted bi-(phosphine oxide), substituted or unsubstitutedtri-(phosphine oxide), substituted or unsubstituted4-(4,6-di(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl)-N,N′-dimethylbenzenamine,where substituted groups R₄₁-R₆₇ are independently selected from, butnot limited to, hydrogen (H), deuterium (D), halogen, direct or branchedalkyl, alkoxy, aryl, alkyl ketone, alkylester, arylester, amide,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl, Each R′ andR″ groups are independently selected from H, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted heteroaryl, halogen, substituted orunsubstituted aryl, substituted or unsubstituted fluoroalkyl,substituted or unsubstituted fluoroaryl, substituted or unsubstitutedalkoxy, substituted or unsubstituted heteroaryl. The alkyl substituentscan include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be 1-50 or higher. L₂ can be bridged by alkyl, aryl, amine and othergroups from R₁-R_(41+n) and to form cryptands which can be coordinatedto Ln to synthesize lanthanide cryptate.

The methods of making the polymers and lanthanide complexes aregenerally well known in the art. FIG. 10G provides an example scheme formaking narrow-band emissive polymers that can include, e.g., generalmonomers as donors and the lanthanide complexes for emission. In themethod shown in FIG. 10G, both the general monomers and the lanthanidecomplexes include amino groups that can be covalently crosslinked withan amine reactive polymer to form lanthanide-complex grafted polymersfor preparing narrow-band emissive polymer dots. As shown, a variety oflanthanide ions can be complexed with the ligands to form the lanthanidecomplex, e.g., Ln can be Ce (III), Pr(III), Nd(III), Sm(III), Eu(III),Tb(III), Dy(III), Ho(III), Er(III), Tm(III), or Yb(III). In someembodiments, the lanthanide ion can be Eu(III) or Tb(III). FIG. 10Hshows another example method of making polymers having lanthanidecomplexes. For example, Ln(TTA)₃Phen-NH₂ and Ln(DPA)₂DPA-NH₂ can beattached to a side chain of a polymer by reaction with the NH₂ groups oneither complex.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude perylene and its derivatives as monomers. Perylene derivativesinclude but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Theperylene derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

FIG. 11A shows example perylene derivatives that can be used as a narrowband monomer in the present invention. The narrow-band monomer can beintegrated into a backbone of the polymer (e.g., by copolymerizing inthe polymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentto R^(1A), R^(1B), R^(2A), R^(2B), R^(2C), R^(2D) or a combinationthereof. As shown by the parentheses in one example, the monomer can beintegrated into the polymer and/or attached by covalent attachment tothe backbone, a terminus, or a sidechain of the polymer by attachment toR^(2B) and R^(2C) groups. In certain embodiments, each of R^(1A),R^(1B), R^(2A), R^(2B), R^(2C) and R^(2D) can be independently selectedfrom the group consisting of, but not limited to, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl, alkyl-substitutedthiophenyl, fluorine (F), cyano (CN) and trifluoro (CF₃). Alkylsubstituted phenyl can include 2-alkylphenyl, 3-alkylphenyl,4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl and 6-alkyl-9,9-dialkyl-substituted fluorenyl.Alkyl-substituted carbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, and 4-alkylthiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃,wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude cyanine and its derivatives as monomers. Cyanine derivativesinclude but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, their extended systems and analogues. Thenarrow-band emissive polymers can also include any other monomers. Thecyanine derivatives can be energy acceptors so that the final Pdots canexhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the above Pdots is less than 70 nm. In certainembodiments, the FWHM is less than 60 nm, less than 50 nm, less than 40nm, less than 30 nm, or less than 20 nm.

FIG. 11B shows one set of example cyanine derivatives that can be usedas a narrow band monomer in the present invention. The narrow-bandmonomer can be integrated into a backbone of the polymer (e.g., bycopolymerizing in the polymer) and/or attached by covalent attachment tothe backbone, a terminus, or a sidechain of the polymer through at leastone attachment to R^(2A), R^(2B), R^(2C), R^(2D), R^(2E), R^(2F),R^(2G), R^(3A), R^(3B), R^(4A), R^(4B), R^(5A), R^(5B), R^(6A), R^(6B)or a combination thereof. In certain embodiments, each of R^(2A),R^(2B), R^(2C), R^(2D), R^(2E), R^(2F), R^(2G), R^(3A) and R^(3B) can beindependently selected from the group consisting of, but not limited to,hydrogen, deuterium, alkyl, aryl, cyano, amino, sulfide, aldehyde,ester, ether, acid, hydroxyl, and halide. R^(4A) and R^(4B) can beindependently selected from the group consisting of, but not limited to,hydrogen, deuterium, alkyl, aryl, acetyl, hydroxyl and phenyl. Incertain embodiments, each of R^(5A) and R^(5B) can be independentlyselected from the group consisting of, but not limited to, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl, and alkyl-substitutedthiophenyl. Alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl and 6-alkyl-9,9-dialkyl-substituted fluorenyl.Alkyl-substituted carbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, and 4-alkylthiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃,wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher. In certain embodiments, each of R^(6A) and R^(6B) can beindependently selected from the group consisting of, but not limited to,alkyl, phenyl, and alkyl-substituted phenyl. Alkyl substituted phenylcan include 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl,2,4-dialkylphenyl, 3,5-dialkylphenyl, and 3,4-dialkylphenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher.

FIG. 11C shows another set of example cyanine derivatives that can beused as a narrow band monomer in the present invention. The narrow-bandmonomer can be integrated into a backbone of the polymer (e.g., bycopolymerizing in the polymer) and/or attached by covalent attachment tothe backbone, a terminus, or a sidechain of the polymer through at leastone attachment to R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B) or acombination thereof. Each of X¹ and X² can include, but is not limitedto, oxygen, sulfur, selenium, and —C(CH₃)₂. In certain embodiments, eachof R^(1A), R^(1B), R^(2A), R^(2B), R³, R^(3A), R^(3B), R⁴, R⁵, R⁶, R⁷,R⁸, R⁹ and R¹⁰ can be independently selected from the group consistingof, but not limited to, hydrogen, deuterium, alkyl, aryl, cyano, amino,sulfide, aldehyde, ester, ether, acid, hydroxyl, and halide. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher.

FIG. 11D shows another set of example cyanine derivatives that can beused as a narrow band monomer in the present invention. The narrow-bandmonomer can be integrated into a backbone of the polymer (e.g., bycopolymerizing in the polymer) and/or attached by covalent attachment tothe backbone, a terminus, or a sidechain of the polymer through at leastone attachment to R^(1A), R^(1B), R^(2A), R^(2B), R^(3A), R^(3B),R^(3C), R^(3D), R⁴, R^(5A), R^(5B), R⁶, or a combination thereof. Eachof X¹ and X² can include, but is not limited to, oxygen, sulfur,selenium, and —C(CH₃)₂. In certain embodiments, each of R^(1A), R^(1B),R^(2A), R^(2B), R^(3A), R^(3B), R^(3C) and R^(3D) can be independentlyselected from the group consisting of, but not limited to, hydrogen,deuterium, alkyl, aryl, cyano, amino, sulfide, aldehyde, ester, ether,acid, hydroxyl, and halide. In certain embodiments, R⁴ can beindependently selected from the group consisting of, but not limited to,hydrogen, deuterium, alkyl, aryl, cyano, amino, sulfide, aldehyde,ester, ether, acid, hydroxyl, and halide. In certain embodiments, eachof R^(5A) and R^(5B) can be independently selected from the groupconsisting of, but not limited to, hydrogen, deuterium, alkyl, cyano,amino, sulfide, aldehyde, ester, ether, acid, hydroxyl, halide, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl, and alkyl-substitutedthiophenyl. Alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl and 6-alkyl-9,9-dialkyl-substituted fluorenyl.Alkyl-substituted carbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, and 4-alkylthiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃,wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher. In certain embodiments, R⁶ can be selected from the groupconsisting of, but not limited to, hydrogen, deuterium, alkyl, cyano,amino, sulfide, aldehyde, ester, ether, acid, hydroxyl, halide, phenyl,alkyl-substituted phenyl, alkyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl, and alkyl-substitutedthiophenyl. Alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl, and3,4-dialkylphenyl. Alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl and 6-alkyl-9,9-dialkyl-substituted fluorenyl.Alkyl-substituted carbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl.Alkyl-substituted thiophenyl can include 2-alkylthiophenyl,3-alkylthiophenyl, and 4-alkylthiophenyl. The alkyl substituents caninclude C_(n)H_(2n+1), or C_(n)F_(2n+1) or —CH₂CH₂[OCH₂CH₂]_(n)—OCH₃,wherein n is 1 to 20. In some embodiments, n can be between 1 to 50 orhigher.

In some embodiments, the narrow-band emissive polymers for making Pdotscan include rhodamine based monomers and their derivatives asnarrow-band monomers. Rhodamine based monomers and their derivativesinclude but are not limited to their alkyl derivatives, arylderivatives, alkyne derivatives, aromatic derivatives, alkoxidederivatives, aza derivatives, rhodamine extended systems and rhodamineanalogues. The narrow-band emissive polymers can also include any othermonomers. The rhodamine based monomers can be energy acceptors so thatthe final Pdots can exhibit narrow-band emissions. The narrow-bandemissive chromophoric polymers in good solvents may exhibit broademissions or narrow emissions. However, their nanoparticle form givesnarrow-band emissions. The emission FWHM of the Pdots includingrhodamine based monomers and their derivatives as narrow-band monomersis less than 70 nm. In certain embodiments, the FWHM can be less than 60nm, less than 50 nm, less than 40 nm, less than 30 nm, or less than 20nm. A variety of other rhodamine derivatives can be used for the presentinvention. In some embodiments, the chromophoric polymer dots of thepresent invention can include a polymer that includes a narrow-bandmonomer having the formula:

wherein each of R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹² andR¹³ can be independently selected from the group consisting of, but notlimited to, hydrogen, deuterium, halogen, cyano, nitro, thiocyanate,isothiocyanate, sulfite, carboxyl, amino, sulfide, aldehyde, ester,ether, acid, linear or branched alkyl, hydroxyl alkyl, aralkyl,alkylene, alkenylene, arylene, heteroarylene, phenylene, azulene,cycloalkylene, alkoxy, aryl, alkyl ketone, alkylester, arylester, amide,fluoroalkyl, fluoroaryl, and polyalkalene (e.g, mehtoxyethoxyethoxy,ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted phenyl, pyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substituted pyridyl,bipyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbipyridyl tripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted tripyridyl, furyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-) substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. The narrow-band monomer can beintegrated into a backbone of the polymer (e.g., by copolymerizing inthe polymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentto R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹² and R¹³ or acombination thereof.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude coumarin based monomers and their derivatives as narrow-bandmonomers. Coumarin based monomers and their derivatives include but arenot limited to their alkyl derivatives, aryl derivatives, alkynederivatives, aromatic derivatives, alkoxide derivatives, azaderivatives, coumarin extended systems and coumarin analogues. Thenarrow-band emissive polymers can also include any other monomers. Thecoumarin based monomers can be energy acceptors so that the final Pdotscan exhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the Pdots including coumarin based monomers andtheir derivatives is less than 70 nm. In certain embodiments, the FWHMis less than 60 nm, less than 50 nm, less than 40 nm, less than 30 nm,or less than 20 nm. A variety of other coumarin derivatives can be usedfor the present invention. In some embodiments, the chromophoric polymerdots of the present invention can include a polymer that includes anarrow-band monomer having the formula:

wherein each of R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ is independently selectedfrom the group consisting of, but not limited to, hydrogen, deuterium,halogen, cyano, nitro, thiocyanate, isothiocyanate, sulfite, carboxyl,amino, sulfide, aldehyde, ester, ether, acid, direct or branched alkyl,hydroxyl alkyl, aralkyl, alkylene, alkenylene, arylene, heteroarylene,phenylene, azulene, cycloalkylene, alkoxy, aryl, alkyl ketone,alkylester, arylester, amide, fluoroalkyl, fluoroaryl, and polyalkalene(e.g, mehtoxyethoxyethoxy, ethoxyethoxy, and —(OCH₂CH₂)_(n)OH, n=1-50),phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedphenyl, pyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted pyridyl, bipyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted bipyridyl tripyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted tripyridyl,furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedfuryl, thienyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted thienyl, pyrrolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted pyrrolyl, pyrazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyrazolyl, oxazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted oxazolyl,thiazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedthiazolyl, imidazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted imidazolyl, pyrazinyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyrazinyl, benzooxadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzooxadizolyl, benzothiadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzothiadizolyl, fluorenyl, alkyl-(alkoxy-,aryl-, fluoroalkyl-, fluoroaryl-)substituted fluorenyl,triphenylaminyl-substituted fluorenyl, diphenylaminyl-substitutedfluorenyl, alkyl-substituted carbazolyl, alkyl-substitutedtriphenylaminyl and alkyl-substituted thiophenyl. As exemplaryembodiments, alkyl substituted phenyl can include 2-alkylphenyl,3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl, 3,5-dialkylphenyl,3,4-dialkylphenyl; alkyl-substituted fluorenyl can include9,9-dialkyl-substituted fluorenyl, 7-alkyl-9,9-dialkyl-substitutedfluorenyl, 6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. The narrow-band monomer can beintegrated into a backbone of the polymer (e.g., by copolymerizing inthe polymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentR¹, R², R³, R⁴, R⁵, and R⁶, or a combination thereof.

In some embodiments, the narrow-band emissive polymers for making Pdotsinclude xanthene based monomers and their derivatives as narrow-bandmonomers. Xanthene based monomers and their derivatives include but arenot limited to their alkyl derivatives, aryl derivatives, alkynederivatives, aromatic derivatives, alkoxide derivatives, azaderivatives, xanthene extended systems and xanthene analogues. Thenarrow-band emissive polymers can also include any other monomers. Thexanthene based monomers can be energy acceptors so that the final Pdotscan exhibit narrow-band emissions. The narrow-band emissive chromophoricpolymers in good solvents may exhibit broad emissions or narrowemissions. However, their nanoparticle form gives narrow-band emissions.The emission FWHM of the Pdots including xanthene based monomers andtheir derivatives is less than 70 nm. In certain embodiments, the FWHMis less than 60 nm, less than 50 nm, less than 40 nm, less than 30 nm,or less than 20 nm. A variety of other xanthene derivatives can be usedfor the present invention. In some embodiments, the chromophoric polymerdots of the present invention can include a polymer that includes anarrow-band monomer having the formula:

wherein each of R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, and R⁹ is independentlyselected from the group consisting of, but not limited to, hydrogen,deuterium, halogen, cyano, nitro, thiocyanate, isothiocyanate, sulfite,carboxyl, amino, sulfide, aldehyde, ester, ether, acid, direct orbranched alkyl, hydroxyl alkyl, aralkyl, alkylene, alkenylene, arylene,heteroarylene, phenylene, azulene, cycloalkylene, alkoxy, aryl, alkylketone, alkylester, arylester, amide, fluoroalkyl, fluoroaryl, andpolyalkalene (e.g, mehtoxyethoxyethoxy, ethoxyethoxy, and—(OCH₂CH₂)_(n)OH, n=1-50), phenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted phenyl, pyridyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted pyridyl, bipyridyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted bipyridyltripyridyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedtripyridyl, furyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted furyl, thienyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-) substituted thienyl, pyrrolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted pyrrolyl,pyrazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazolyl, oxazolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted oxazolyl, thiazolyl, alkyl-(alkoxy-, aryl-,fluoroalkyl-, fluoroaryl-)substituted thiazolyl, imidazolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substituted imidazolyl,pyrazinyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedpyrazinyl, benzooxadizolyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted benzooxadizolyl, benzothiadizolyl,alkyl-(alkoxy-, aryl-, fluoroalkyl-, fluoroaryl-)substitutedbenzothiadizolyl, fluorenyl, alkyl-(alkoxy-, aryl-, fluoroalkyl-,fluoroaryl-)substituted fluorenyl, triphenylaminyl-substitutedfluorenyl, diphenylaminyl-substituted fluorenyl, alkyl-substitutedcarbazolyl, alkyl-substituted triphenylaminyl and alkyl-substitutedthiophenyl. As exemplary embodiments, alkyl substituted phenyl caninclude 2-alkylphenyl, 3-alkylphenyl, 4-alkylphenyl, 2,4-dialkylphenyl,3,5-dialkylphenyl, 3,4-dialkylphenyl; alkyl-substituted fluorenyl caninclude 9,9-dialkyl-substituted fluorenyl,7-alkyl-9,9-dialkyl-substituted fluorenyl,6-alkyl-9,9-dialkyl-substituted fluorenyl,7-triphenylaminyl-9,9-dialkyl-substituted fluorenyl and7-diphenylaminyl-9,9-dialkyl-substituted fluorenyl; alkyl-substitutedcarbazolyl can include N-alkyl-substituted carbazolyl,6-alkyl-substituted carbazolyl and 7-alkyl-substituted carbazolyl;alkyl-substituted triphenylaminyl can include 4′-alkyl-substitutedtriphenylaminyl, 3′-alkyl-substituted triphenylaminyl,3′,4′-dialkyl-substituted triphenylaminyl and 4′,4″-alkyl-substitutedtriphenylaminyl; alkyl-substituted thiophenyl can include2-alkylthiophenyl, 3-alkylthiophenyl, and 4-alkylthiophenyl. The alkylsubstituents can include C_(n)H_(2n+1), or C_(n)F_(2n+1) or—CH₂CH₂[OCH₂CH₂]_(n)—OCH₃, wherein n is 1 to 20. In some embodiments, ncan be between 1 to 50 or higher. The narrow-band monomer can beintegrated into a backbone of the polymer (e.g., by copolymerizing inthe polymer) and/or attached by covalent attachment to the backbone, aterminus, or a sidechain of the polymer through at least one attachmentto R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, and R⁹ or a combination thereof.

In some embodiments, these narrow-band emissive monomers can beincorporated into the backbone of the conventional semiconductingpolymer to obtain narrow-band emissive polymers. In this embodiment, thenarrow-band emissive monomers can be copolymerized with other monomerssuch as fluorene monomer, phenylene vinylene monomer, phenylene monomer,benzothiadiazole monomer, thiophene monomer, carbazole monomer, and anyother monomers to form narrow-band emissive polymers. In someembodiments, the narrow-band emissive units can be chemically linked tothe side chains of the conventional semiconducting polymer to obtainnarrow-band emissive polymers. In this embodiment, conventionalluminescent semiconducting polymers include, but are not limited tofluorene polymers, phenylene vinylene polymers, phenylene polymers,benzothiadiazole polymers, thiophene polymers, carbazole fluorenepolymers and their copolymers, and any other conventional semiconductingpolymers.

In some embodiments, the ratio of the narrow-band monomers in thebackbone (or the narrow-band emissive units in the side-chains) relativeto other monomers in the polymer can be adjusted to obtain narrow-bandemissive Pdots. As provided further herein, the ratios can be describeda variety of ways that will be understood by one of ordinary skill inthe art. For example, monomers in a polymer of the present invention canbe identified as x and 1−x or x, y and z in which x+y+z=1.Alternatively, the ratios can be described as X and Y, in which theratio can be X/(X+Y). In another way, the ratio can be identified withrespect to the number of monomers (n) in a polymer (e.g., one monomercan be 0.02n, a second monomer can be 0.45n, a third monomer can be0.03n, and a fourth monomer can be 0.5n). In some embodiments, the ratioof the narrow-band emissive monomer to other monomer is 1:1000. In someembodiments, the ratio of the narrow-band emissive monomer to othermonomer is 1:500. In some embodiments, the ratio of the narrow-bandemissive monomer to other monomer is 1:100. In some embodiments, theratio of the narrow-band emissive monomer to other monomer is 1:90. Insome embodiments, the ratio of the narrow-band emissive monomer to othermonomer is 1:80. In some embodiments, the ratio of the narrow-bandemissive monomer to other monomer is 1:70. In some embodiments, theratio of the narrow-band emissive monomer to other monomer is 1:60. Insome embodiments, the ratio of the narrow-band emissive monomer to othermonomer is 1:50. In some embodiments, the ratio of the narrow-bandemissive monomer to other monomer is 1:45. In some embodiments, theratio of the narrow-band emissive monomer to other monomer is 1:40. Insome embodiments, the ratio of the narrow-band emissive monomer to othermonomer is 1:35. In some embodiments, the ratio of the narrow-bandemissive monomer to other monomer is 1:30. In some embodiments, theratio of the narrow-band emissive monomer to other monomer is 1:25. Insome embodiments, the ratio of the narrow-band emissive monomer to othermonomer is 1:20. In some embodiments, the ratio of the narrow-bandemissive monomer to other monomer is 1:15. In some embodiments, theratio of the narrow-band emissive monomer to other monomer is 1:10, 1:9,1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 1:0.5, 1:0.1, or higher. Avariety of ratios can be optimized to obtain narrow-band emissiondepending on the monomers used. In certain embodiments, the ratio of thenarrow-band monomer to the general monomer is less than about 1:1, lessthan about 0.8:1, less than about 0.6:1, less than about 0.5:1, lessthan about 0.4:1, less than about 0.3:1, less than about 0.2:1, lessthan about 0.1:1, less than about 0.08:1, less than about 0.06:1, lessthan about 0.04:1, or less than about 0.02:1. Example 1 and FIG. 13describe an example approach to obtain narrow-band emission by adjustingthe concentration of narrow-band monomers.

In some embodiments, the narrow-band emissive polymers (i.e. polymerscomprising narrow-band monomers) may have narrow-band emissions in agood solvent, such as some hydrophobic polymer in tetrahydrofuransolution. After forming these polymers into Pdot nanoparticles in water,the Pdot also exhibit narrow-band emission. In this embodiment, thenanoparticle formation does not cause obvious change of the emissionbandwidth. In some embodiments, the FWHM of the Pdot emission in wateris less than about 70 nm. In some embodiments, the FWHM of the Pdotemission in water is less than about 60 nm. In some embodiments, theFWHM of the Pdot emission in water is less than about 50 nm. In someembodiments, the FWHM of the Pdot emission in water is less than about45 nm. In some embodiments, the FWHM of the Pdot emission in water isless than about 40 nm. In some embodiments, the FWHM is less than about35 nm, 30 nm, 25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18 nm, 17nm, 16 nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less.

In some embodiments, the narrow-band emissive polymers (i.e. polymerscomprising narrow-band monomers) may exhibit broad-band emissions in agood solvent, such as some hydrophobic polymer in tetrahydrofuransolution. However, after forming these polymers into Pdot nanoparticlesin water, the Pdots exhibit narrow-band emissions. In a good solvent,hydrophobic semiconducting polymers typically adopt an extended rod-likeconformation, and the inter-chain energy transfer is not efficient. Whenthe polymers are densely packed into a compact nanoparticle, becauseintra-particle energy transfer and inter-chain energy transfer are muchmore efficient in the nanoparticle form, therefore the resulting Pdotshave narrow-band emissions. Example 3 and 4, FIGS. 15 and 16 describeexamples of synthesis and design of such polymers and Pdots. In someembodiments, the FWHM of the Pdot emission in water is less than about70 nm. In some embodiments, the FWHM of the Pdot emission in water isless than about 60 nm. In some embodiments, the FWHM of the Pdotemission in water is less than about 50 nm. In some embodiments, theFWHM of the Pdot emission in water is less than about 45 nm. In someembodiments, the FWHM of the Pdot emission in water is less than about40 nm. In some embodiments, the FWHM of the Pdot emission is less thanabout 35 nm, 30 nm, 25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18nm, 17 nm, 16 nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less.

In some embodiments, the narrow-band emissive polymers (i.e. polymerscomprising narrow-band monomers) may have narrow-band emissions in agood solvent, such as some hydrophobic polymer in toluene solution.After forming these polymers into Pdot nanoparticles in water usingnanoprecipitation, however, the Pdots exhibit broad-band emissionsbecause of the complex backbone folding behaviors, disorderedmorphologies and chain aggregation. In this embodiment, the narrow-bandPdots may be prepared using the miniemulsion method that may maintainthe narrow emission from the polymer. In some embodiments, the emissionFWHM of the Pdot formed by miniemulsion is less than about 70 nm. Insome embodiments, the emission FWHM in water is less than about 60 nm.In some embodiments, the emission FWHM in water is less than about 50nm. In some embodiments, the emission FWHM in water is less than about45 nm. In some embodiments, the emission FWHM in water is less thanabout 40 nm. In some embodiments, the emission FWHM in water is lessthan about 35 nm, 30 nm, 25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19nm, 18 nm, 17 nm, 16 nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, orless.

In some embodiments, the narrow-band emissive chromophoric polymer dotscan include narrow-band emissive polymers physically blended orchemically cross-linked with conventional broad-band semiconductingpolymers. In some embodiments, the conventional semiconducting polymermay have energy transfer to the narrow-band emissive polymers so thatthe final Pdots have narrow-band emissions. In some embodiments, thenarrow-band emissive polymers can be chemically cross-linked with two ormore broad-band polymers (FIG. 31A). The broad-band polymers can beenergy donors and the narrow-band emissive polymer can be energyacceptors. Multi-step energy transfer occurs from the broad-band polymerto the narrow-band emissive polymer so that the polymer dots givenarrow-band emissions. The chemical cross-linking between polymers canuse the functional reactive groups such as haloformyl, hydroxyl,aldehyde, alkenyl, alkynyl, anhydride, carboxamide, amines, azocompound, carbonate, carboxylate, carboxyl, cyanates, ester, haloalkane,imine, isocyanates, nitrile, nitro, phosphino, phosphate, phosphate,pyridyl, sulfonyl, sulfonic acid, sulfoxide, thiol groups. Thesefunctional groups can be attached to the side chains and/or the terminusof each polymer chain.

The concentration of the narrow-band emissive polymers relative tobroad-band semiconducting polymers can be adjusted to maximize thefluorescence performance of the narrow-band emissive Pdots, such asnarrow emission FWHM, high fluorescence quantum yield, desirablefluorescence lifetime etc. In some embodiments, the FWHM of the Pdotemission is less than about 70 nm. In some embodiments, the FWHM of thePdot emission is less than about 60 nm. In some embodiments, the FWHM ofthe Pdot emission is less than about 50 nm. In some embodiments, theFWHM of the Pdot emission is less than about 45 nm. In some embodiments,the FWHM of the Pdot emission is less than about 40 nm. In someembodiments, the FWHM of the Pdot emission is less than about 35 nm, 30nm, 25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18 nm, 17 nm, 16nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less.

In some embodiments, the narrow-band emissive chromophoric polymer dotscan include conventional broad-band semiconducting polymers chemicallycross-linked with other narrow-band emissive species. The narrow-bandemissive species include but are not limited to small organic dyemolecules, metal complexes, metal clusters, lanthanide complexes. Insome embodiments, the semiconducting polymer may have energy transfer tothe narrow-band emissive species so that the final Pdots havenarrow-band emissions. The concentration of the narrow-band emissivespecies relative to semiconducting polymers can be adjusted to maximizethe fluorescence performance of the narrow-band emissive Pdots, such asnarrow emission FWHM, high fluorescence quantum yield, desirablefluorescence lifetime etc. In some embodiments, the FWHM of the Pdotemission is less than about 70 nm. In some embodiments, the FWHM of thePdot emission is less than about 60 nm. In some embodiments, the FWHM ofthe Pdot emission is less than about 50 nm. In some embodiments, theFWHM of the Pdot emission is less than about 45 nm. In some embodiments,the FWHM of the Pdot emission is less than about 40 nm. In someembodiments, the FWHM of the Pdot emission is less than about 35 nm, 30nm, 25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18 nm, 17 nm, 16nm, 15 nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less.

In a particular embodiment, the narrow-band emissive chromophoricpolymer dots can include polymer dots embedded with inorganic quantumdots. In some embodiments, the semiconducting polymer may have energytransfer to the quantum dots so that the final composite Pdots havenarrow-band emissions. Example 9 and FIGS. 26-28 describe the Pdotsembedded with inorganic quantum dots to obtain narrow-band emission. Theconcentration of the quantum dots relative to semiconducting polymerscan be adjusted to maximize the fluorescence performance of thenarrow-band emissive Pdots, such as narrow emission FWHM, highfluorescence quantum yield, etc. In some embodiments, the FWHM of thecomposite Pdot emission is less than about 70 nm. In some embodiments,the FWHM of the composite Pdot emission is less than about 60 nm. Insome embodiments, the FWHM of the composite Pdot emission is less thanabout 50 nm. In some embodiments, the FWHM of the composite Pdotemission is less than about 45 nm. In some embodiments, the FWHM of thecomposite Pdot emission is less than about 40 nm. In some embodiments,the FWHM of the composite Pdot emission is less than about 35 nm, 30 nm,25 nm, 24 nm, 23 nm, 22 nm, 21 nm, 20 nm, 19 nm, 18 nm, 17 nm, 16 nm, 15nm, 14 nm, 13 nm, 12 nm, 11 nm, 10 nm, or less.

In some embodiments, the narrow-band emissive chromophoric polymer dotincludes narrow-band emissive chromophoric polymer chemicallycross-linked with small organic dye molecules, metal complexes, and anycombinations thereof. These dyes or metal complexes may have sensingfunctions, such as oxygen sensing capability, ion sensing capability,glucose sensing capability, neurotransmitter sensing capability, drugsensing capability, metabolite sensing capability, protein sensingcapability, signaling molecule sensing capability, toxin sensingcapability, DNA and RNA sensing capability, and the like.

In some embodiments, the narrow-band emissive chromophoric polymer dotmay also include narrow-band emissive semiconducting polymer, physicallymixed or chemically cross-linked with other components including, e.g.inorganic luminescent materials, to tune emission color, improve quantumyield and photostability, and the like.

Functionalization and Bioconjugates of Chromophoric Polymers Dots withNarrow-band Emissions

In some embodiments, this invention provides narrow-band emissive Pdotsfunctionalized with a functional group. As used herein the term“functional group” refers to any chemical unit that can be attached,such as by any stable physical or chemical association, to thechromophoric polymer dot, thereby rendering the surface of thechromophoric polymer dot available for conjugation or bioconjugation. Insome embodiments, functional groups can be hydrophobic functionalgroups. Examples of hydrophobic functional groups include but notlimited to alkyne, strained alkyne, azide, diene, alkene, cyclooctyne,and phosphine groups (for click chemistry). In some embodiments,functional groups can be hydrophilic functional groups. Examples ofhydrophilic functional groups include but not limited to carboxylic acidor salts thereof, amino, mercapto, azido, diazo, aldehyde, ester,hydroxyl, carbonyl, sulfate, sulfonate, phosphate, cyanate, succinimidylester, substituted derivatives thereof. Such functional groups can befound by one of ordinary skill in the art, for example in BioconjugateTechniques (Academic Press, New York, 1996 or later versions) thecontent of which is herein incorporated by reference in its entirety forall purposes.

In some embodiments, a functional group can be created with covalentbonding to the backbone, side chain, or terminating unit of thenarrow-band emissive chromophoric polymer. Therefore, the resultingpolymer dots exhibit narrow-band emission and simultaneously havefunctional groups for bioconjugation. Such functional groups could befound by one of ordinary skill in the art, for example in BioconjugateTechniques (Academic Press, New York, 1996 or later versions) thecontent of which is herein incorporated by reference in its entirety forall purposes. In some embodiments, each narrow-band emissive polymer dotmay have only one functional group. In some embodiments, eachnarrow-band emissive polymer dot may have only two functional groups.The two functional groups can be the same or different. In someembodiments, each narrow-band emissive polymer dot may have only threeor more functional groups. The three or more functional groups can bethe same or different.

In some embodiments, the present invention provides a bioconjugatecomprising a narrow-band emissive chromophoric polymer dot as describedabove and a biomolecule, wherein the biomolecule is attached to thepolymer dot either directly or indirectly by the functional group. Thebioconjugates also include narrow-band emissive chromophoric polymer dotas described above, associated with biological particle such as virus,bacteria, cells, biological or synthetic vesicles such as liposomes. Theterm “biomolecule” is used to describe a synthetic or naturallyoccurring protein, glycoprotein, peptide, amino acid, metabolite, drug,toxin, nuclear acid, nucleotide, carbohydrate, sugar, lipid, fatty acidand the like. Desirably, the biomolecule is attached to the functionalgroup of narrow-band emissive chromophoric polymer dot via a covalentbond. For example, if the functional group of the polymer dot is acarboxyl group, a protein biomolecule can be directly attached to thepolymer dot by cross-linking the carboxyl group with an amine group ofthe protein molecule. In some embodiments, each narrow-band emissivepolymer dot may have only one biomolecule attached. In some embodiments,each narrow-band emissive polymer dot may have only two biomoleculeattached. The two biomolecules can be the same or different. In someembodiments, each narrow-band emissive polymer dot may have only threeor more biomolecules attached. The three or more biomolecules can be thesame or different. In some embodiments, the biomolecular conjugationdoes not change substantively the emissive properties of the narrow-bandemissive Pdots. For example, the bioconjugation does not broaden theemission spectra, does not reduce fluorescence quantum yield, does notchange the photostability etc.

Methods for Preparing Chromophoric Polymers Dots with Narrow-BandEmissions

A variety of polymerization reactions can be used for synthesis of thepolymers described herein. For example, semiconducting polymersincluding homo-polymer and multi-component copolymer or heteropolymercan be synthesized by using a variety of different reactions.Non-limiting examples of reactions for synthesizing semiconductingpolymers include the Heck, Mcmurray and Knoevenagel, Wittig, Horner,Suzuki-Miyaura, Sonogashira, Yamamoto and Stille coupling reaction andso on. Other polymerization strategies such as electropolymerization,oxidative polymerization can also be employed to make semiconductingpolymers. Furthermore, microwave-assisted polymerization takes less timeand often can give higher molecular weight and yield.

Several of the above mentioned polymerization reactions are shown below,e.g., by using synthesis of polyfluorene and its derivatives asexamples. The examples below show homo-polymer, but the reactions forthe synthesis of heteropolymer or copolymer is similar except that thestarting monomers are different. Using these reactions, we can formvarious polymers from the individual units or monomers described in thisinvention. The monomers and any of the substituents on the monomers(such as the substituents described herein) can also be made usingstandard synthesis methods generally well known in the art.

1) Stille polycondensation for synthesis of conjugated polymers

2) Suzuki polycondensation for synthesis of conjugated polymers.

3) Yamamoto polycondensation for synthesis of conjugated polymers.

4) Sonogashira polycondensation for synthesis of conjugated polymers.

5) Heck reaction for synthesis of conjugated polymers.

6) Mcmurray and Knoevenagel reaction for synthesis of conjugatedpolymers.

7) Mcmurray and Knoevenagel reaction for synthesis of conjugatedpolymers.

8) Electropolymerization for synthesis of conjugated polymers.

9) Wittig reaction for Synthesis of conjugated polymers.

10) Horner-Wadsworth-Emmons reaction for synthesis of conjugatedpolymers.

11) Oxidative polymerization for synthesis of conjugated polymers.

In some embodiments, narrow-band emissive chromophoric polymer dot canbe prepared by using the solvent mixing method. The solvent mixingmethod involves quick mixing a solution of the chromophoric polymer in agood solvent (such as tetrahydrofuran) with a miscible solvent (such aswater) to fold the polymer into a nanoparticle form, and Pdots can beobtained after removal of the good solvent. In some embodiments, thenarrow-band emissive polymer dots can also be prepared by an emulsion orminiemulsion method, based on shearing a mixture comprising twoimmiscible liquid phases (such as water and another immiscible organicsolvent) with the presence of a surfactant.

In one aspect, the present invention can include methods of making apolymer dot. The methods can include providing a solvent solutioncomprising a chromophoric polymer, the chromophoric polymer being in anelongated coil form and comprising a narrow-band monomer, and whereinthe chromophoric polymer includes an emission spectrum with a full widthhalf maximum (FWHM) of less than about 70 nm; and mixing the solventsolution comprising the chromophoric polymer with a miscible solvent toform a condensed chromophoric polymer, wherein the condensedchromophoric polymer includes an emission spectrum with a FWHM less thanabout 70 nm. In another aspect, the present invention can include amethod of making a polymer dot that includes providing a solventsolution comprising a chromophoric polymer, the chromophoric polymerbeing in an elongated coil form and comprising a narrow-band monomer,and wherein the chromophoric polymer includes an emission spectrum witha full width half maximum (FWHM) of greater than about 70 nm; and mixingthe solvent solution comprising the chromophoric polymer with a misciblesolvent to form a condensed chromophoric polymer, wherein the condensedchromophoric polymer includes an emission spectrum with a FWHM less thanabout 70 nm.

In some embodiments, polymer dots can be made as condensed polymernanoparticles that have intrachain energy transfer between, e.g., anarrow-band monomer and one or more general monomers on the same polymerchain. The present invention can further include methods of makingpolymer dots by physically blending and/or chemically crosslinking twoor more polymer chains together. For example, the polymer dots can haveinterchain energy transfer in which a condensed polymer nanoparticle caninclude two or more polymer chains physically blended and/or chemicallycrosslinked together. For interchain energy transfer, one of the chainsmay include a narrow-band monomer and another chain may include one ormore general monomers that can act as an energy donor to the narrow bandmonomer, which is an energy acceptor. As provided in the methods ofmaking described above, broad-band polymer chains in good solvent (e.g.,polymers that have a FWHM of greater than 70 nm) can be condensed andalso physically blended and/or crosslinked so as to produce a narrowband emitting polymer dot (e.g., a polymer dot that has a FWHM of lessthan 70 nm). Some of the polymer dots can be made to have bothintrachain and interchain energy transfer. In some instances, thecombination of intrachain and interchain energy transfer can increasethe quantum yield of the polymer dots. In certain embodiments, the finalPdots can exhibit narrow-band emissions because of energy transfer tothe narrow-band monomers.

Methods of Using Chromophoric Polymer Dots with Narrow-Band Emissions

The present invention further provides methods of using the narrow-bandemissive polymer dots described herein. For example, the presentinvention provides methods of fluorescence-based detection using thenarrow-band emissive polymer dots as a novel class of fluorescent probeand their bioconjugates for a variety of applications, including but notlimited to flow cytometry, fluorescence activated sorting,immunofluorescence, immunohistochemistry, fluorescence multiplexing,single molecule imaging, single particle tracking, protein folding,protein rotational dynamics, DNA and gene analysis, protein analysis,metabolite analysis, lipid analysis, FRET based sensors, high throughputscreening, cell detection, bacteria detection, virus detection,biomarker detection, cellular imaging, in vivo imaging, bioorthogonallabeling, click reactions, fluorescence-based biological assays such asimmunoassays and enzyme-based assays, and a variety of fluorescencetechniques inbiological assays and measurements. In certain aspects, thepolymer dots disclosed herein can be used for methods of detection thatinvolve multiplexing over a variety of wavelength ranges.

In one aspect, the present invention provides methods for imagingpolymer dots that include administering a population of polymer dotsdescribed herein to a subject and exciting at least one polymer dot inthe population of polymer dots, e.g., with an imaging system. The methodcan further include detecting a signal from at least one excited polymerdot in the population of polymer dots. As described further herein, thepolymer dots can be administered in a composition.

In another aspect, the present invention includes a method of multiplexdetection with a polymer dot. The method can include detecting thepolymer dot with a detector system comprising a filter configured topass a spectrum of light having a full width half maximum (FWHM) of lessthan about 70 nm, wherein the polymer dot includes a condensedchromophoric polymer comprising a narrow-band monomer, the condensedchromophoric polymer having an emission spectrum that is substantiallypassed through the filter. In certain embodiments, the FWHM is less thanabout 60 nm, less than about 50 nm, less than about 40 nm, less thanabout 30 nm, or less than about 20 nm. As described further herein, thepolymer dots of the present invention can include, e.g., a homopolymeror heteropolymer including a narrow band monomer, such as BODIPY and/orBODIPY derivative monomer, a squaraine and/or squaraine derivative, ametal complex and/or metal complex derivative monomer, a porphyrinand/or porphyrin derivative monomer, a phthalocyanine and/orphthalocynanine derivative monomer, a lanthanide complex and/orlanthanide complex derivative monomer, a perylene and/or perylenederivative monomer, a cyanine and/or cyanine derivative monomer, arhodamine and/or rhodamine derivative monomer, a coumarin and/orcoumarin derivative monomer, and/or a xanthene and/or xanthenederivative monomer. A narrow band unit can be, e.g., a narrow bandmonomer or a fluorescent nanoparticle embedded in or attached to thepolymer dot. The fluorescent nanoparticle can be, e.g., a quantum dot. Anarrow band unit can also include a polymer or fluorescent dye moleculethat gives a narrow emission in a polymer dot of the present invention.

The present invention also provides methods and compositions foradministering the polymer dots described herein to a subject tofacilitate diagnostic and/or therapeutic applications. In one aspect,the present invention provides a method for administering a polymer dotcomposition. The method can include administering a polymer dotcomposition described herein to a subject. A subject can include, but isnot limited to, a mouse, a rat, a rabbit, a human, or other animal. Incertain embodiments, the compositions can include a population ofpolymer dots and a pharmaceutically acceptable excipient. Pharmaceuticalexcipients useful in the present invention include, but are not limitedto, binders, fillers, disintegrants, lubricants, coatings, sweeteners,flavors and colors. One of skill in the art will recognize that otherpharmaceutical excipients are useful in the present invention.

The polymer dots of the present invention can be administered asfrequently as necessary, including hourly, daily, weekly or monthly. Thecompounds utilized in the methods of the invention can be, e.g.,administered at dosages ranging from, for example, about 1 mg to about510 mg, or about 0.0125 mg/kg body weight to about 6.375 mg/kg bodyweight (assuming an average adult weighs 80 kg). The dosages, however,may be varied depending upon the requirements of the subject theseverity of the condition being treated and/or imaged, and/or thepolymer dot being employed. For example, dosages can be empiricallydetermined considering the type and stage of disease diagnosed in aparticular patient and/or the type of imaging modality being used inconjunction with the polymer dots. The dose administered to a subject,in the context of the present invention should be sufficient to effect abeneficial diagnostic or therapeutic response in the subject. The sizeof the dose also can be determined by the existence, nature, and extentof any adverse side-effects that accompany the administration of aparticular polymer dot in a particular subject. Determination of theproper dosage for a particular situation is within the skill of thepractitioner.

The compositions described herein can be administered to the patient ina variety of ways, including parenterally, intravenously, intradermally,intramuscularly, colonically, rectally or intraperitoneally. In someembodiments, the pharmaceutical compositions can be administeredparenterally, intravenously, intramuscularly or orally. The oral agentscomprising a population of the polymer dots of the invention can be inany suitable form for oral administration, such as liquid, tablets,capsules, or the like. The oral formulations can be further coated ortreated to prevent or reduce dissolution in stomach.

The polymer dot compositions of the present invention can beadministered to a subject using any suitable methods known in the art.Suitable formulations for use in the present invention and methods ofdelivery are generally well known in the art. For example, a populationof polymer dots described herein can be formulated as pharmaceuticalcompositions with a pharmaceutically acceptable diluent, carrier orexcipient. A population of polymer dots of the present invention can beadministered in any pharmaceutically acceptable composition.

Furthermore, a population of polymer dots can be formulated forparenteral, topical, nasal, sublingual, gavage, or local administration.For example, the pharmaceutical compositions are administeredparenterally, e.g., intravenously, subcutaneously, intradermally, orintramuscularly, or intranasally. Thus, the invention providescompositions for parenteral administration that include a solution of asingle or mixture of a population of polymer dots described herein,dissolved or suspended in an acceptable carrier, e.g., an aqueouscarrier. The compositions may contain pharmaceutically acceptableauxiliary substances as required to approximate physiological conditionsincluding pH adjusting and buffering agents, tonicity adjusting agents,wetting agents and the like, such as, for example, sodium acetate,sodium lactate, sodium chloride, potassium chloride, calcium chloride,sorbitan monolaurate, triethanolamine oleate, etc.

The present invention also provides kits for administering the polymerdots to a subject for treating and/or diagnosing a disease state. Suchkits typically include two or more components useful for administration.Components can include polymer dots of the present invention, reagents,containers and/or equipment.

In certain embodiments, the kits of the present invention can includepackaging assemblies that can include one or more components. Forexample, a packaging assembly may include a container that houses atleast one of the polymer dot compositions as described herein. Aseparate container may include other excipients or agents that can bemixed with the polymer dot compositions prior to administration to apatient. In some embodiments, a physician may select and match certaincomponents and/or packaging assemblies depending on the particulardiagnostic and/or therapeutic application.

The specific dimensions of any of the apparatuses, devices, systems, andcomponents thereof, of the present invention can be readily varieddepending upon the intended application, as will be apparent to those ofskill in the art in view of the disclosure herein. Moreover, it isunderstood that the examples and embodiments described herein are forillustrative purposes only and that various modifications or changes inlight thereof may be suggested to persons skilled in the art and areincluded within the spirit and purview of this application and scope ofthe appended claims. Numerous different combinations of embodimentsdescribed herein are possible, and such combinations are considered partof the present invention. In addition, all features discussed inconnection with any one embodiment herein can be readily adapted for usein other embodiments herein. The use of different terms or referencenumerals for similar features in different embodiments does notnecessarily imply differences other than those expressly set forth.Accordingly, the present invention is intended to be described solely byreference to the appended claims, and not limited to the preferredembodiments disclosed herein.

EXAMPLES

The following examples are included to further describe some aspects ofthe present invention, and should not be used to limit the scope of theinvention.

Example 1 Synthesis of BODIPY Monomer2a (FIG. 12) and a Series ofFluorene-BODIPY Copolymers at Different BODIPY Concentrations

The present example provides a method for obtaining narrow-band BODIPYmonomer2a and a series of fluorene-BODIPY copolymers at different BODIPYconcentrations.

Instrumentation and characterizations for Synthesis. ¹H (500 MHz), ¹³C(125 MHz) NMR spectra were recorded on Bruker AV500 spectrometers. ¹HNMR and ¹³C NMR spectra used tetramethylsilane (TMS) as an internalstandard in CDCl₃. Mn and molecular weight distribution [weight-averagemolecular weight/number-average molecular weight (Mw/Mn)] values of allpolymers were estimated by size exclusion chromatography (SEC) with aTOSOH G3000HXI system equipped with three consecutive polystyrene gelcolumns [TOSOH gels: ¦Á-4000, ¦Á-3000, and ¦Á-2500] and refractive-indexand ultraviolet detector at 40° C. The system was operated at a flowrate of 1.0 mL/min with a tetrahydrofuran as an eluent. Polystyrenestandards were employed for calibration. All the chemicals werepurchased from Sigma-Aldrich and TCI America.

Synthesis of BODIPY Monomer 2a(8-Mesityl-1,3,5,7-tetramethyl-4,4-difluoro-4-bora-3a,4a-diaza-s-indacene).110 μl of trifluoroacetic acid in dry CH₂Cl₂ (10 ml) was added slowly toa solution of 2,4,6-trimethylbenzaldehyde (1.482 g, 10 mmol) and2,4-dimethyl-1H-pyrrole (2.38 g, 25 mmol) in dry CH₂Cl₂ (250 ml) at roomtemperature. 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (2.27 g, 10 mmol)is added after 3 h stirring under ice bath cooling and stirred for 20min. The solution is stirred for an additional 1 h at room temperature.NEt₃ (20 mL, 144 mmol) is added, followed by slow addition of BF₃. Et₂O(23 ml, 170 mmol). The reaction mixture is washed after 12 h of stirringat room temperature with saturated aqueous Na₂CO₃ solution (2×150 ml),dried over Na₂SO₄, and concentrated on a rotary evaporator. The brown,oily residue is purified by column chromatography on silica withhexane/CH₂Cl₂=3:1. The product fraction with greenish fluorescence isdried to yield a red-brown solid. Yield: 2.3 g, 62.8%. ¹H NMR (500 MHz,CDCl₃): δ=6.979 (s, 2H), 5.993 (s, 2H), 2.592 (s, 6H), 2.368 (s, 3H),2.128 (s, 6H), 1.417 (s, 6H). ¹³C NMR (125MHz, CDCl₃): δ=155.09, 142.31,141.68, 138.57, 134.92, 131.13, 130.62, 129.0, 120.79, 21.22, 19.51,14.64, 13.41.

Synthesis of fluorene-BODIPY copolymer series. BODIPY fluorene copolymerseries with different BODIPY monomer molar ratio (2%, 5%, 10%, 25%, 50%)are synthesized by palladium-catalyzed Suzuki coupling reaction from9,9-dioctylfluorene and BODIPY monomer. 9,9-Dioctyl-2,7-dibromofluorene,9,9-Dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester,BODIPY monomer 1a, 2 drops of aliquot 336, 10 ml of 2M Na₂CO₃ aqueoussolution, 15 ml of toluene were placed in a 50 ml flask. The flask wasevacuated and refilled with N₂ four times by using the freeze/thawmethod and Pd(PPh₃)₄ (1-1.5 mol %) was added. The flask was furtherdegassed four times, then reaction was heated to 80° C. and stirredunder N₂. After 70 h 0.2 ml of bromobenzene and 15 mg of phenylboronicacid were added to end-cap the polymer chain and the reaction wasstirred for an additional 2 h at 80° C. respectively. The whole mixturewas poured into 200 ml of MeOH, filtered, and washed with 0.2M of HCl.The precipitate was stirred in 50 ml of acetone at room temperature for24 h and dried in vacuum oven to obtain dark pink to dark red solid.Yield: 73-81%. NMR results: for PFO-BODIPY10 ¹H NMR (500 MHz, CDCl₃):δ=7.89-7.61 (m), 7.53 (m), 7.42 (m, 6H), 7.25 (m, 5H), 7.05 (m, 2H),2.69 (s, 6H), 2.39 (s, 3H), 2.32 (s, 6H), 2.09-2.17 (s, 4H), 1.31 (s,6H), 1.19 (s, 24), 0.87 (s, 6H). ¹³C NMR (125MHz, CDCl₃): δ=154.07,151.85, 151.74, 151.08, 141.94, 140.55, 140.08, 138.16, 135.05, 133.96,132.24, 132.17130.63, 129.15, 128.96, 128.83, 128.57, 128.47, 127.25,126.82, 126.19, 124.86, 121.53, 120.01, 119.55, 55.39, 55.29, 40.44,30.08, 29.76, 29.26, 29.19, 23.95, 22.64, 21.31, 19.91, 14.11, 13.65,11.74. Mn: 23048, Mw: 43610, PDI: 1.89.

Example 2 Synthesis of BODIPY Monomer1a (FIG. 12) and a Narrow-bandEmissive Fluorene-BODIPY Copolymer Polymer510

The present example provides a method for obtaining narrow-band BODIPYmonomer2a and a narrow-band emissive fluorene-BODIPY copolymerpolymer510.

Synthesis of 4-methyl-3,5-diiodobenzaldehyde for BODIPY monomer1a.Powdered I₂ (3.04 g, 12 mmol) and then NaIO₄ (0.86 g, 4 mmol) were addedslowly to stirred 98% H₂SO₄ (50 ml). Stirring was continued for 30 minat room temperature to give a dark brown iodinating solution.p-Tolualdehyde (1.5 g, 14 mmol) was added in one portion to theiodinating solution and the resulting solution was stirred overnight atroom temperature. Then the reaction mixture was slowly poured intostirred ice water. The crude solid products were collected byfiltration, washed with water until the filtrates were neutral, vacuumdried in the dark to get light brown powder, and re-crystallized fromethyl acetate to give light yellow solid. Yield: 2.13 g, 40.9%. ¹H NMR(CDCl₃, 500MHz): δ=9.823 (s, 1H), 8.306 (d, 2H), 2.842 (s, 3H). ¹³C NMR(CDCl₃, 125MHz): δ=189.19, 162.98, 150.32, 140.83, 99.97, 35.98.

Synthesis of BODIPY Monomer1a. To a solution of4-methyl-3,5-diiodobenzaldehyde (1.5 g, 4.2 mmol) and2,4-dimethyl-1H-pyrrole (1 g, 10.5 mmol) in dry CH₂Cl₂ (120 ml) is addeda solution of 110 μl trifluoroacetic acid in dry CH₂Cl₂ (5 ml) slowly atroom temperature. 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (0.95 g, 4.2mmol) is added after 3 h stirring under ice bath cooling and stirred for10 min. The solution is stirred for an additional 1 h at roomtemperature. NEt₃ (10 ml, 72 mmol) is added, followed by slow additionof BF₃. Et₂O (12 mL, 81 mmol). The reaction mixture is washed after 10 hof stirring at room temperature with saturated aqueous Na₂CO₃ solution(2×100 ml), dried over Na₂SO₄, and concentrated on a rotary evaporator.The brown, oily residue is purified by column chromatography on silicawith hexane/CH₂Cl₂=3:1. The product fraction with greenish fluorescenceis dried to yield an orange solid. Yield: 0.48 g, 19.5%. ¹H NMR (CDCl₃,500MHz): δ=7.831 (s, 2H), 6.042 (s, 2H), 2.874 (s, 3H), 2.581 (s, 6H),1.544 (s, 6H). ¹³C NMR (CDCl₃, 125MHz): δ=156.25, 144.12, 142.83,138.94, 135.89, 131.11, 121.67, 99.09, 34.93, 15.14, 14.61.

Synthesis of monomer4 (FIG. 21) for Polymer510. A mixture of2,7-dibromofluorene (15 mmol, 4.86 g), tert-butyl 3-bromopropanoate (33mmol, 6.86 g), sodium hydroxide solution (40%, 35 mL), Bu₄NBr (1.5 mmol,0.48 g), toluene (70 mL) was stirred at 85° C. overnight. The organicphase was separated, washed with water and dried over MgSO₄. Afterevaporation of the solvent, the residue was purified by columnchromatography (DCM). The product was obtained as a white solid. Yield:4.81 g, 83%. ¹HNMR (500 MHz, CDCl₃): δ=7.47-7.54 (m, 6H), 2.30 (t, 4H),1.47 (t, 4H), 1.33 (s, 18H). ¹³CNMR (125 MHz, CDCl₃): 172.71, 150.47,139.60, 131.56, 126.99, 122.57, 121.93, 80.97, 54.58, 34.92, 30.36,28.52.

Synthesis of Polymer510. In the glovebox under nitrogen atmosphere, adry three neck 50 mL round-bottom flask with stir bar was charged with248 mg (0.9 mmol) of bis(1,5-cyclooctadiene) nickel(0), 97.1 mg (0.9mmol) of cyclooctadiene, and 140.6 mg (0.9 mmol) of bypyridine in 9.0 mLof a 1:1 mixture of toluene and dimethylformamide (DMF). A dark purplecolor then developed. The solution was heated to 60° C. In the glovebox,a dry 20 mL flask was charged with 7.04 mg (0.008 mmol) of BODIPYmonomer 1a, 241.5 mg (0.376 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and9.7 mg (0.016 mmol) of monomer 4 in 4.0 mL of a 1:1 mixture of tolueneand DMF, then they were added dropwise into the above catalyst mixture.The flask containing this solution was covered with foil to protect itfrom light and the reaction mixture was refluxed for 4 days. 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 6 h at 60° C. The product was precipitated in30 mL of a 1:1 mixture of methanol and concentrated hydrochloric acid.The polymer was dissolved in dichloromethane and washed with aqueous 15wt % of sodium thiosulfate solution (3×30 mL) followed by washing withMilli-Q water and drying over MgSO₄, for the removal of residual iodinefrom polymer. The concentrated polymer solution in dichloromethane waspoured into 100 ml of MeOH, filtered. The precipitate was stirred in 50ml of acetone at room temperature for 24 h. The polymer was obtained asa yellow powder. Deprotection of the tert-butyl esters was then followedby adding 1 ml of trifluoroacetic acid into a solution of polymer (70mg) in DCM (40 ml) and stirred overnight. The organic layer was washedwith water (100 ml×4) and concentrated to 10 ml and precipitated inmethanol (100 ml). The final powder was collected by filtration, washedwith acetone, and dried in vacuum oven to obtain green solid. yield: 101mg, 64.5%. 1H NMR (CDCl₃, 500 MHz): δ=7.89-7.61 (m), 7.53 (m), 7.42 (m,6H), 2.09-2.16 (s, 4H), 1.18 (s, 24H), 0.85 (s, 6H). ¹³C NMR (CDCl₃, 125MHz): δ=151.86, 140.56, 140.06, 126.20, 121,55, 120.00, 55.38, 40.43,31.84, 30.08, 29.27, 23.97, 22.64, 14.11.

Example 3 Synthesis of BODIPY Monomer2a (FIG. 12) and a Narrow-bandEmissive Fluorene-BODIPY Copolymer Polymer590

The present example provides a method for obtaining narrow-band BODIPYmonomer2a and a narrow-band emissive fluorene-BODIPY copolymerpolymer590.

Synthesis of BODIPY monomer 2a. A 250 ml round-bottom flask was firstcharged with 2.2 g (6 mmol) of BODIPY monomer dissolved in 80 ml ofethanol. To this solution 4.57 g (18 mmol) of powdered I₂ was added andallowed to dissolve. 2.15 g (12.2 mmol) of HIO₃ was dissolved in a 0.7 gof water, and this solution was added dropwise by a syringe over a 20min. After the addition was complete, the solution was heated to 60° C.and refluxed for 5 h. Ethanol was removed on a rotary evaporator. Theresidue was purified by column chromatography with a silica withhexane/CH₂Cl₂=3:1. The product 2a was dried to obtain a metallic darkred solid. Yield: 2.5 g, 68%. ¹H NMR (500MHz, CDCl₃): δ=7.008 (d, 2H),2.682 (s, 6H), 2.391 (s, 3H), 2.096 (s, 6H), 1.437 (s, 6H). ¹³C NMR (125MHz, CDCl₃): δ=156.42, 144.57, 141.72, 139.29, 134.81, 130.86, 130.52,129.31, 85.30, 21.28, 19.55, 16.06, 15.80.

Synthesis of Polymer590. Fluorene-BODIPY copolymer Polymer590 aresynthesized by palladium-catalyzed Suzuki coupling reaction from9,9-dioctylfluorene, benzo[c]-1,2,5-thiadiazole and BODIPY monomer.4,7-Dibromobenzo[c]-1,2,5-thiadiazole (52.9 mg, 0.18 mmol),9,9-Dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester(111.68 mg, 0.20 mmol), BODIPY monomer 2a (5.03 mg, 0.008 mmol), monomer4 (7.3 mg, 0.012 mmol), 2 drops of aliquat 336, 10 ml of 2M Na₂CO₃aqueous solution, 15 ml of toluene were placed in a 50 ml round bottomflask. The flask was evacuated and refilled with N₂ four times by usingthe freeze/thaw method and Pd(PPh₃)₄ (10 mg, 0.0086 mmol) was added. Theflask was further degassed four times, then reaction was heated to 80°C. and stirred under N₂. After 70 h 0.2 ml of bromobenzene and 15 mg ofphenylboronic acid were added to end-cap the polymer chain and thereaction was stirred for an additional 2 h at 80° C. respectively. Thewhole mixture was poured into 200 ml of MeOH, filtered, and washed with0.2M of HCl. The dried polymer was stirred in 50 ml of acetone at roomtemperature for 24 h. Polymer was obtained as a brown powder.Deprotection of the tert-butyl esters was then followed by adding 1 mlof trifluoroacetic acid into a solution of polymer in DCM (40 ml) andstirred overnight. The organic layer was washed with water (150 ml×5)and concentrated to 10 ml and precipitated in methanol (100 ml). Thefinal powder was collected by filtration, washed with acetone, and driedin vacuum oven to get a brown solid. Yield: 112 mg, 73.2%. ¹H NMR(500MHz, CDCl₃): δ=8.14-8.08 (m,2 H), 8.01-7.72 (m), 7.83-7.73 (m),7.40-7.43 (m, 6H), 7.24 (m, 5H), 7.09 (m, 2H), 6.97 (m,2H), 3.95 (s,4H),2.53 (s, 6H), 2.40 (s, 6H), 2.19 (s, 8H), 1.51 (m, 6 H), 1.20 (s, 24 H),0.85 (m, 6H). ¹³C NMR (125 MHz, CDCl₃): δ=154.43, 151.84, 140.95,136.52, 133.67, 128.39, 128.03, 124.06, 120.12, 55.5, 55.27, 40.3,31.89, 31.79, 30.17, 29.33, 29.31, 24.11, 22.67, 14.13. Mn: 14480, Mw:28396, PDI: 1.96.

Example 4 Synthesis of BODIPY Monomer3a (FIG. 12) and a Narrow-bandEmissive Fluorene-BODIPY Copolymer Polymer680

The present example provides a method for obtaining narrow-band BODIPYmonomer2a and a narrow-band emissive fluorene-BODIPY copolymerpolymer590.

Synthesis of BODIPY Monomer 3a. p-tolualdehyde (392 mg, 4.24 mmol),monomer 2a (500 mg, 0.81 mmol), p-toluene sulfonic acid (90 mg),piperidine (3 ml) were dissolved in 100 ml of benzene reflux for 12 h byusing Dean-Stark. The mixture was cooled to room temperature, thesolvents were removed under vacuum, and the crude product was purifiedby column chromatography on silica gel eluted with ethyl acetate/hexane1:7. The crude was recrystallized from chloroform/methanol to give theproduct as a metallic shiny solid. Yield: 420 mg, 62.3%. ¹H NMR (500MHz,CDCl₃): δ=8.157-8.191 (s, 2H), 7.689-7.722 (s, 2H), 7.589-7.605 (s, 4H),7.258-7.274 (s, 4H), 7.029 (s, 2H), 2.435 (s, 6H), 2.409 (s, 3H), 2.127(s, 6H), 1.512 (s 6H). ¹³C NMR (125 MHz, CDCl₃): δ=150.41, 145.17,139.50, 139.48, 139.35, 139.32, 135.27, 134.05, 132.11, 131.32, 129.57,129.33, 127.71, 117.98, 82.62, 21.53, 21.31, 19.73, 16.28.

Synthesis of Polymer680.4,7-Bis(2-bromo-5-thienyl)-2,1,3-benzothiadiazole (5.5 mg, 0.012 mmol),4,7-Dibromobenzo[c]-1,2,5-thiadiazole (49.4 mg, 0.168 mmol),9,9-Dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester(111.68 mg, 0.20 mmol), BODIPY monomer 3a (6.58 mg, 0.008 mmol), monomer4 (7.3 mg, 0.012 mmol), 2 drops of aliquat 336, 10 ml of 2M Na₂CO₃aqueous solution, 15 ml of toluene were placed in a 50 ml round bottomflask. The flask was evacuated and refilled with N₂ four times by usingthe freeze/thaw method and Pd(PPh₃)₄ (10 mg, 0.0086 mmol) was added. Theflask was further degassed four times, then reaction was heated to 80°C. and stirred under N₂. After 70 h 0.2 ml of bromobenzene and 15 mg ofphenylboronic acid were added to end-cap the polymer chain and thereaction was stirred for an additional 2 h at 80° C. respectively. Thewhole mixture was poured into 300 ml of MeOH, filtered, and washed with0.2M of HCl. The dried precipitate was stirred in 50 ml of acetone atroom temperature for 24 h. Polymer 1b was obtained as a dark brownpowder. Deprotection of the tert-butyl esters was then followed byadding 1 ml of trifluoroacetic acid into a solution of polymer in DCM(40 ml) and stirred overnight. The organic layer was washed with water(150 ml×5) and concentrated to 10 ml and precipitated in methanol (100ml). The final powder was collected by filtration, washed with acetone,and dried in vacuum oven to obtain a dark brown solid. Yield: 70 mg,62.1%. ¹H NMR (500MHz, CDCl₃): δ=8.23 (m, 2H), 8.08-8.14 (m, 2H),8.02-7.98 (m, 2H), 7.85-7.83 (m, 2H), 7.78 (m, 2H), 7.58 (m, 4H),7.44-7.38 (m, 4H), 7.21 (m, 4H), 7.08 (m, 2H), 6.97 (m, 2H), 2.38 (s,3H), 2.33 (s, 6H), 1.48 (s, 6H), 1.20 (s, 24 H), 0.85 (m, 6H). ¹³C NMR(125 MHz, CDCl₃): δ=154.42, 151.82, 140.95, 136.53, 133.66, 128.37,128.03, 124.08, 120.11, 55.49, 55.26, 40.28, 31.88, 30.17, 30.12, 29.33,29.30, 24.11, 22.66, 14.11.

Example 5 Preparation and Characterizations of Narrow-band EmissivePdots by Using Fluorene-BODIPY Copolymers

The present example provides the preparation and characterizations ofnarrow-band emissive Pdots by using fluorene-BODIPY copolymers.

Fluorene-BODIPY Pdots were prepared by nanoprecipitaiton. A solution ofpolymer precursor in THF (2 mL, 100 ppm) was quickly injected into water(10 mL) under ultrasonication. THF was evaporated by N₂ flow at 70° C.and the solution was concentrated to 4-5 mL, followed by filtrationthrough a 0.2 micron filter. The particle size and zeta-potentials ofPdots in bulk solution was characterized by dynamic light scattering(Malvern Zetasizer NanoS). TEM measurements were recorded on atransmission electron microscope (FEI Tecnai F20). UV-Vis absorptionspectra were recorded with a DU 720 scanning spectrophotometer (BeckmanCoulter, Inc., CA USA) using 1 cm quartz cuvettes. Fluorescence spectrawere obtained using a commercial Fluorolog-3 fluorometer (HORIBA JobinYvon, N.J. USA). Fluorescence quantum yields were measured using aHamamatsu photonic multichannel analyzer C10027 equipped with CCDintegrating sphere.

FIG. 13A shows a series of fluorene-BODIPY copolymers that include ageneral monomer fluorene and a narrow-band monomer (BODIPY Monomer 2a inFIG. 12) at different molar ratios. FIG. 13B shows the fluorescencespectra of the polymers in a good solvent tetrahydrofuran (THF). FIG.13C shows the fluorescence spectra of the Pdots in water (THF). As canbe seen, the emission spectra of the polymers in THF exhibit similarFWHM for all the polymers. However, the Pdots shows quite different FWHMbecause of the densely packing of polymer chromophores in thenanoparticles. Narrow-band emissions can be obtained by adjusting theBODIPY ratio relative to the general fluorene monomer. FIG. 14A showsthe chemical structure of a narrow-band emissive polymer (Polymer510)synthesized by using the BODIPY monomer1a as narrow-band monomer andseveral general monomers. FIG. 14B shows the absorption spectrum andemission spectra of the Polymer510 Pdots in water. When excited at 380nm, the Pdot emission exhibits a FWHM of 41 nm. When excited at 470 nm,the Pdot emission exhibits a FWHM of 25 nm. The fluorescence quantumyield was measured to be 64%. These properties indicate that narrow-bandemission can be obtained in the Pdots.

FIG. 15A shows the chemical structure of a narrow-band emissive polymer(Polymer2 590) synthesized by using the BODIPY monomer 2a as narrow-bandmonomer and several general monomers. FIG. 15B shows the absorption andemission spectra of the Polymer590 in THF. FIG. 15C shows the absorptionand emission spectra of the Polymer590 Pdots in water. The Pdot emissionexhibits a FWHM of 64 nm and fluorescence quantum yield is 0.13. Asindicated by the spectra, the polymer shows broad emission in a goodsolvent such as THF. However, the Pdots show narrow-band emission. FIG.16A shows the chemical structure of a narrow-band emissive polymer(Polymer3 680) synthesized by using the BODIPY monomer 3a as narrow-bandmonomer and several general monomers. FIG. 16B shows the absorption andemission spectra of the Polymer680 in THF. FIG. 16C shows the absorptionand emission spectra of the Polymer680 Pdots in water. The Pdot emissionexhibits a FWHM of 55 nm and fluorescence quantum yield is 0.19. Asindicated by the spectra, the polymer shows broad emission in a goodsolvent such as THF. However, the Pdots show narrow-band emission. FIG.17 show the particle size distributions of narrow-band emissivePolymer510 Pdots, Polymer590 Pdots, and Polymer680 Pdots, respectively.The data were measured by dynamic light scattering. All the Pdots showsmall particle size in the range of 10˜20 nm.

Example 6 Bioconjugation of Narrow-band Emissive Fluorene-BODIPY Pdots

The present example provides a method for bioconjugation to thenarrow-band emissive fluorene-BODIPY Pdots.

Bionconjugation was performed by utilizing the EDC-catalyzed reactionbetween carboxyl groups on fluorene-BODIPY Pdots surface and aminegroups on biomolecules. In a typical bioconjugation reaction, 80 μL ofpolyethylene glycol (5% w/v PEG, MW 3350) and 80 μL of concentratedHEPES buffer (1 M) were added to 4 mL of functionalized Pdot solution(50 μg/mL in MilliQ water), resulting in a Pdot solution in 20 mM HEPESbuffer with a pH of 7.3. Then, 240 μL of streptavidin (purchased fromInvitrogen (Eugene, Oreg., USA)) was added to the solution and mixedwell on a vortex. 80 μL of freshly-prepared EDC solution (10 mg/mL inMilliQ water) was added to the solution, and the above mixture was lefton a rotary shaker. After 4 hours at room temperature, Triton-X 100(0.25% (w/v), 80 μL) and BSA (2% (w/v), 80 μL) were added. The mixturewas then left on rotary shaker for one hour. Finally, the resulting Pdotbioconjugates were separated from free biomolecules by gel filtrationusing Sephacryl HR-300 gel media.

Example 7 Flow Cytometry and Confocal Imaging of MCF-7 Cells Labeledwith Narrow-band Emissive Fluorene-BODIPY Pdots

The present example provides a method for using the narrow-band emissivefluorene-BODIPY Pdots for cellular labeling.

Cell Culture. The breast cancer cell line MCF-7 was ordered fromAmerican Type Culture Collection (ATCC, Manassas, Va., USA). Cells werecultured at 37° C., 5% CO₂ in Eagles minimum essential mediumsupplemented with 10% Fetal Bovine Serum (FBS), 50 U/mL penicillin, and50 μg/mL streptomycin. The cells were cultured prior to experimentsuntil confluence was reached. The cells were harvested from the cultureflask by briefly rinsing with culture media followed by incubation with5 mL of Trypsin-EDTA solution (0.25 w/v % Trypsin, 0.53 mM EDTA) at 37°C. for 5-15 minutes. After complete detachment, the cells were rinsed,centrifuged, and resuspended in 1×PBS buffer. The cell concentration wasdetermined by microscopy using a hemocytometer.

Specific labeling for flow cytometry. For specific cell labeling withthe narrow-band emissive Pdot-streptavidin (Pdot-SA), a million cellswere blocked with BlockAid blocking buffer (Invitrogen, Eugene, Oreg.,USA) and then were incubated sequentially with biotinylated primaryanti-EpCAM antibody (used to label the cell-surface EpCAM receptors onMCF-7 cells) and 20 μg/mL (based on Pdots) Pdot-SA for 30 minutes each,followed by two washing steps using labeling buffer. Finally, thespecifically labeled cells were fixed in 0.6 mL 4% (v/v)paraformaldehyde solution. For the control labeling, no biotinylatedprimary anti-EpCAM antibody was added. Flow cytometry was operated on aBD FACS Canto flow cytometer (BD Biosciences, San Jose, Calif., USA). A488 nm laser was used for excitation and emission was collected throughFITC channel equipped with a 500 nm longpass filter and a 530/30 nmband-pass filter. Data was analyzed using the FACSDiva1 software. FIG.18 shows flow cytometry results of Polymer590 Pdots and Polymer680 Pdotslabeled MCF-7 cells, respectively. Panel A shows the side scattering(SSC) versus forward scattering (FSC). Panel B shows the fluorescenceintensity distributions of the MCF-7 cells labeled with Polymer590Pdots. Blue curve is the negative control, and Orange curve is thepositive labeling. Panel C shows the fluorescence intensitydistributions of the MCF-7 cells labeled with Polymer680 Pdots. Greencurve is the negative control, and Red curve is the positive labeling.As can be seen, both the Polymer590 and Polymer680 Pdots are specificfor labeling the cellular targets without nonspecific labeling.

Specific labeling for cellular surface imaging. For labeling cellularsurface with the narrow-band emissive Pdot-SA conjugates, live MCF-7cells in the glass-bottomed culture dish were blocked with BlockAidblocking buffer (Invitrogen, Eugene, Oreg., USA). Then the MCF-7 cellswere incubated sequentially with biotinylated primary anti-EpCAMantibody (were used to label the cell-surface EpCAM receptors on MCF-7cells) and 5 nM Pdot-SA for 30 minutes each, followed by two washingsteps after each incubation. For the control, no biotinylated primaryanti-EpCAM antibody was added. The Pdot-tagged cells were thencounterstained with Hoechst 34580 and imaged immediately on afluorescence confocal microscope (Zeiss LSM 510). FIGS. 19A and 19Bshows fluorescence images of MCF-7 breast-cancer cells labeled withPolymer590 Pdot-streptavidin and Polymer590 Pdot-streptavidin,respectively. Negative labeling performed under the same condition butin the absence of the biotinylated primary antibody does not showfluorescence signal, again indicating the narrow-band emissive Pdotprobes are specific for cellular labeling.

Example 8 Synthesis of Narrow-band Emissive Polymers (e.g., PFS andPFS5.5) Using Squaraine Derivatives as Narrow-band Monomers and theOptical Characterizations

The present example provides example methods for obtaining a narrow-bandemissive fluorene-squaraine copolymers, such as PFS (polymer690) (shownin FIG. 24A) and PFS5.5 (shown in FIG. 24B).

Synthesis of Fluorene-squaraine Copolymer PFS (Polymer690). Toluene (8ml) and Na₂CO₃ (2M, 5 ml) were in the flask and degassed for half hour.A mixture of 9,9-dioctyl-2,7-dibromofluorene (105 mg, 0.192 mmol),9,9-dioctyl-2,7-dibromofluorene (112 mg, 0.2 mmol),2,5-bis[(5-bromo-1-hexadecyl-3,3-dimethyl-2,3-dihydroindole-2-ylidene)methyl]cyclobutendiylium-1,3-diolate(8 mg, 0.008 mmol), Bu₄NBr (3 mg, 0.008 mmol) were added. The mixturewas degassed and refilled with N₂ (repeated 4 times) before addition ofPd(PPh₃)₄ (8 mg, 0.007 mmol), the resulting mixture was stirred at 90°C. for 40 hours and phenylboronic acid (20 mg) dissolved in THF (0.2 ml)was added. After 2 hour, bromobenzene (0.2 ml) was added and thenstirred for more 3 hours. The mixture was poured into MeOH (40 ml), thenwas filtered the precipitate. The precipitate were washed by MeOH, H₂Oand Acetone to remove monomers, small oligomer and salts. The solid wasdissolved in DCM (5 ml), filtered through 0.2 um membrane andre-precipitated in MeOH (30 ml), the solid was stirred in acetone (40ml) for 4 Hours and filtered off and dried in high vacuum.

FIG. 24A shows multi-step synthesis of narrow-band emissive polymer PFS(Polymer690) using a squaraine derivative as narrow-band monomer andfluorene as general monomer. FIG. 25A shows photophysical data of thenarrow-band emissive fluorene-4% squaraine copolymer and fluorene-19%squaraine copolymer. FIG. 25B shows absorption spectra data of thefluorene-4% squaraine copolymer and fluorene-19% squaraine copolymer.FIG. 25C shows emission spectra of the fluorene-4% squaraine copolymerand fluorene-19% squaraine copolymer excited at 405 nm. The fluorene-4%squaraine copolymer Pdots shows emission FWHM of 37 nm at 690 nm andfluorescence quantum yield is 0.23. FIG. 25D shows emission spectra ofthe fluorene-4% squaraine copolymer and fluorene-19% squaraine copolymerexcited at 675 nm. These data shows that both narrow-band emission andhigh fluorescence quantum yield can be obtained in thefluorene-squaraine copolymer based Pdots.

Synthesis of 5-Bromo-2,3,3-trimethylindolenine (Compound 2). A mixtureof 4-bromophenylhydrazine (4.46 g, 20 mmol), isopropylmethylketone (3.44g, 40 mmol), EtOH (80 mL) and concentrated H₂SO₄ (1.86 g, 40 mmol) in a125 mL round bottom flask was heated under reflux for overnight. Aftercooling, the mixture was diluted with CH₂Cl₂ (100 mL) and was washedwith 10% NaHCO₃ (100 ml) twice and water (100 mL) twice, then dried overMagnesium sulfate and filtered. The solution was then passed through ashort column quickly, and evaporated under reduced pressure to get 4.25g as a reddish oil. (Yield: 90%). ¹H NMR (500 MHz, CDCl₃)=7.40˜7.38 (m,3H), 2.25 (s, 3H), 1.28 (s, 6H). ¹³C NMR (CDCl₃)=187.8, 152.2, 147.38,130.1, 124.3, 120.8, 118.3, 53.5, 22.4, 14.9. HRMS (ESI): (M⁺,C₁₁H₁₂BrN) calcd 237.0153; found 237.0150.

Synthesis of 5-Bromo-1-hexadecyl-2,3,3-trimethyl-3H-indolium Iodide(Compound 3). A mixture of 5-Bromo-2,3,3-trimethylindolenine 2 (900 mg,3.78 mmol), 1-iodohexadecane (1.6 g, 4.45 mmol) and nitromethane (5 mL)was refluxed for overnight. After cooling and concentrating the mixtureunder reduced pressure, diethyl ether (25 mL) was added. The solutionwas cooled to 4° C. for 1 h, and the precipitate was collected, thenwashed with diethyl ether (50 mL) and dried. The yellow solid, 1.5 g,was obtained (Yield: 70%). ¹H NMR (500 MHz, CDCl₃)=7.70˜7.62 (m, 3H),4.63 (t, J=7.6 Hz, 2H), 3.08 (s, 3H), 1.91˜1.87 (m, 2H), 1.66 (s, 6H),1.43˜1.21 (m, 26H), 0.85 (t, J=7.0, 3H). ¹³C NMR (CDCl₃)=195.6, 143.4,139.9, 132.6, 126.6, 124.3, 117.0, 54.7, 50.4, 31.7, 29.5, 29.4, 29.3,29.1, 28.9, 27.7, 26.6, 23.0, 22.5, 17.2, 13.9. HRMS (ESI): (M⁺,C₂₇H₄₅BrIN) calcd 589.1780; found 589.1782.

Synthesis of 5-bromo-1-hexadecyl-3,3-dimethyl-2-methyleneindoline(Compound 4). 5-Bromo-1-hexadecyl-2,3,3-trimethyl-3H-indolium Iodide 3(2.92 g, 3.26 mmol) was suspended in 2N NaOH aqueous solution (50 mL)and diethyl ether (50 mL), stirred for 30 minutes, extracted withdiethyl ether and water, then dried and evaporated under vacuum. Theproduct was a yellowish oil, 1.84 g (Yield: 98%). ¹H NMR (500 MHz,CDCl₃)=7.26 (s, 1H), 7.20 (d, J=2.0 Hz, 1H), 7.13 (s, 1H), 3.85 (d,J=2.0 Hz, 2H), 2.43 (t, J=7.3 Hz, 2H), 1.61 (m, 2H), 1.30˜1.22 (m, 26H),0.88 (t, J=7.0 Hz, 3H). HRMS (ESI): (M⁺, C₂₇H₄₄BrN) calcd 461.2657;found 461.2661.

Synthesis of2,5-Bis[(5-bromo-1-hexadecyl-3,3-dimethyl-2,3-dihydroindole-2-ylidene)methyl]cyclobutendiylium(Compound 1). A mixture of 3,4-dihydroxy-3-cyclobutene-1,2-dione 4 (105mg, 0.9 mmol) and5-Bromo-1-hexadecyl-3,3-dimethyl-2-methylene-2,3-dihydroindole (840 mg,1.84 mmol) in toluene/butanol (1:1, 15 mL) was refluxed overnight with aDean-Stark trap. After cooling to room temperature, the solvent wasremoved under vacuum. The residue was purified by silica gel withchromatography (PE/EA), and the product was obtained as a dark greensolid, 500 mg (Yield: 50%). ¹H NMR (500 MHz, CDCl₃)=7.45˜7.41 (m, 4H),6.87 (d, J=8.2Hz, 2H), 6.61 (s, 2H), 3.98 (s, 4H), 1.81˜1.75 (m, 16H),1.44˜1.28(m, 52H), 0.86 (t, J=6.5 Hz, 6H). ¹³C NMR (CDCl₃)=182.2, 180.1,169.4, 141.5, 130.6, 125.6, 116.5, 110.6, 49.3, 43.8, 31.9, 29.7, 29.67,29.65, 29.57, 9.50, 29.44, 29.33, 27.02, 26.95, 22.67, 14.1.

General procedure for the snthesis of squaraine polymer PFS. Polymer PFSwas synthesized using different feeding ratios of monomer by SuzukiCoupling. For example, in the synthesis of 1.5% PFS,9,9-Doctylfluorene-2,7-diboronic acid bis(1,3-propanediol)ether (11.68mg, 0.2 mmol), 9,9-Dioctyl-2,7-dibromofluorene (105.3 mg, 0.193 mmol),2,5-Bis[(5-bromo-1-hexadecyl-3,3-dimethyl-2,3-dihydroindole-2-ylidene)methyl]cyclobutendiylium1 (8 mg, 0.008 mmol), TBAB (2.5 mg, 0.008 mmol), and Pd(PPh₃)₄ (8 mg,3.5 mol %) was added in Toluene/2N Na₂CO₃ 1:1 (10 mL). The mixture wasdegassed and refilled with N₂, then refluxed for 2 days. Phenylboronicacid (20 mg), dissolved in THF (0.5 mL), was added, and after 2 h,bromobenzene (0.5 mL) was added and further stirred for 3 h. The mixturewas poured into Methanol (100 mL), and the precipitate was filtered,washed with methanol, water and acetone to remove monomer, smalloligomers, and inorganic salts. The crude product was dissolved in DCM(7 mL), filtered with 0.2 μm membrane and re-precipitated in methanol(75 mL). The powder was stirred in acetone (100 mL) for 4 h andcollected by filtration and dried in vacuum.

Synthesis of 6-bromo-2,3,3-trimethyl-3H-benzo[f]indole (Compound 7).6-bromonaphthalen-2-amine (2.5 g, 11.25 mmol) was used to synthesizecompound 6, ³ (2.1 g, yield 80%). Without purification and following theprocedure for making compound 2, compound 6 was used to synthesizecompound 7 to obtain 2 g of product as a reddish oil (Yield: 80%). ¹HNMR (500 MHz, CDCl₃)=8.09 (s, 1H), 7.88 (d, J=8.9 Hz, 2H), 7.79˜7.74 (m,2H), 7.61˜7.59 (m, 1H), 2.37 (s, 3H), 1.52 (s, 6H). ¹³C NMR(CDCl₃)=151.2, 149.2, 133.3, 132.3, 131.9, 131.6, 131.3, 131.1, 129.6,128.6, 127.9, 127.0, 124.1, 124.0, 121.0, 118.1, 23.1, 22.2, 15.7. HRMS(ESI): (M⁺, C₁₅H₁₄BrN) calculated 287.0310; found 287.0308.

Synthesis of 6-bromo-1-hexadecyl-2,3,3-trimethyl-3H-benzo[f]indol-1-iumiodide (Compound 8). Following the procedure to make compound 3, weobtained compound 9 as a yellow oil, 1.5 g (Yield: 70%). ¹H NMR (500MHz, CDCl₃)=8.17 (s, 1H), 8.00˜7.95 (m, 2H), 7.83˜7.77 (m, 2H), 4.76 (t,J=7.6 Hz, 2H), 3.17 (s, 3H), 1.95˜1.86 (m, 2H), 1.45˜1.17 (m, 32H), 0.85(t, J=7.0, 3H). HRMS (ESI): (M⁺, C₃₁H₄₇BrIN) calcd 639.1937; found639.1940.

Synthesis of6-bromo-1-hexadecyl-3,3-dimethyl-2-methylene-2,3-dihydro-1H-benzo[f]indole(Compound 9). Following the procedure to make compound 4, we obtainedcompound 9 as a yellowish oil, 1.88 g (Yield: 90%). ¹H NMR (500 MHz,CDCl₃)=7.87 (s, 1H), 7.80 (d, J=9.1 Hz, 1H), 7.57 (d, J=8.7 Hz, 1H),7.42˜7.39(m, 1H), 6.95 (d, J=8.7 Hz, 1H), 3.94˜3.92 (m, 2H), 3.56˜3.54(m, 2H), 1.65 (m, 2H), 1.33˜1.20 (m, 26H), 0.87 (t, J=7.0 Hz, 3H). HRMS(ESI): (M⁺, C31H46BrN) calcd 511.2814; found 511.2816.

Synthesis of6-bromo-2-((Z)-(3-((E)-(6-bromo-1-hexadecyl-3,3-dimethyl-1H-benzo[f]indol-2(3H)-ylidene)methyl)-2-hydroxy-4-oxocyclobut-2-en-1-ylidene)methyl)-1-hexadecyl-3,3-dimethyl-3H-benzo[f]indol-1-ium(Compound 5)

Following procedure to make compound 4, we obtained compound 5 as a darkgreen solid, 350 mg (Yield: 45%). ¹H NMR (500 MHz, CDCl₃)=8.06˜8.04 (m,4H), 7.77 (d, J=8.7 Hz, 2H), 7.64˜7.62 (m, 2H), 7.30˜7.29 (m, 2H), 6.02(s, 2H), 4.08 (s, 4H), 1.86˜1.61 (m, 16H), 1.45˜1.21 (m, 52H), 0.87 (t,J=6.6 Hz, 6H). ¹³C NMR (CDCl₃)=182.7, 178.1, 170.9, 139.8, 134.4, 133.1,132.8, 132.1, 131.5, 131.2, 131.1, 130.9, 130.4, 130.0, 129.5, 129.2,128.6, 127.9, 126.9, 125.7, 124.7, 124.1, 123.8, 123.2, 117.7, 111.4,110.7, 87.1, 86.0, 77.3, 77.0, 76.7, 50.9, 44.6, 43.7, 42.7, 32.6, 31.7,30.8, 30.3, 29.5, 29.4, 29.2, 28.5, 28.2, 27.7, 27.2, 26.9, 26.6, 26.2,25.3, 23.5, 22.5, 21.8, 21.6, 14.4, 14.2, 13.6.

General procedure for the synthesis of squaraine polymer PFS 5.5.Polymer PFS5.5 was synthesized using different feeding ratios of monomerby Suzuki Coupling following the same procedure as for the synthesis ofthe PFS polymer.

Preparation of Pdots. A solution of polymer PFS or PFS 5.5 with 20 wt %of PS-PEG-COOH or PSMA in THF (4 mL, 50 ppm) was injected into water (10mL) under ultrasonication, respectively. THF was evaporated by N₂ flowat 70° C. and the solution was concentrated to 4-5 mL, followed byfiltration through a 0.2 micron filter. Bionconjugation was performed byutilizing the EDC-catalyzed reaction between carboxyl groups on Pdots'surface and amine groups on biomolecules. In an example bioconjugationreaction, 80 μL of polyethylene glycol (5% w/v PEG, MW 3350) and 80 μLof concentrated HEPES buffer (1 M) were added to 4 mL of functionalizedPdot solution (50 mg/mL in MilliQ water), resulting in a Pdot solutionin 20 mM HEPES buffer with a pH of 7.3. Then, 240 μL of streptavidin(from Invitrogen (Eugene, Oreg., USA)) was added to the solution andmixed well on a vortex. 80 μL of freshly-prepared EDC solution (10 mg/mLin MilliQ water) was added to the solution, and the above mixture wasleft on a rotary shaker. After 4 hours at room temperature, Triton-X 100(0.25% (w/v), 80 μL) and BSA (2% (w/v), 80 μL) were added. The mixturewas then left on rotary shaker for one hour. Finally, the resulting Pdotbioconjugates were separated from free biomolecules by gel filtrationusing Sephacryl HR-300 gel media.

Single-particle brightness measurement. For the measurement ofsingle-particle fluorescence brightness, samples were diluted in Milli-Qwater, dried on cleaned glass coverslips (previously functionalized with(3-aminopropyl)trimethoxysilane (APTMS)), and imaged on a customizedhome-built wide-field epifluorescence microscope. Fluorescence intensityemitted per frame for a given particle was estimated by integrating theCCD signal over the fluorescent spot. The wide-field microscope wasconstructed by directing the 488-nm laser beam from a Sapphire laser(Coherent, Santa Clara, Calif. USA) or 405 nm laser beam from a diodelaser (World Star Technologies, Toronto, Canada) into an invertedmicroscope (Nikon TE2000U, Melville, N.Y., USA) using home-builtsteering optics. Laser excitation power was measured at the nosepiecebefore the objective. The objective used for illumination and lightcollection was a Nikon CFI Plan Fluor 100XS Oil (with iris) objectivewith 100× magnification and 0.5-1.3 N.A (Nikon, Melville, N.Y., USA).Fluorescence signal was filtered by a 500 nm long pass filter (HQ500LP;Chroma, Rockingham, Vt., USA) and imaged onto an EMCCD camera(Photometrics Cascade: 512B, Tucson, Ariz. USA).

Spectroscopic characterizations and biological applications. Squarainebased monomers were synthesized and narrow-band emissive polymer dotsbased on fluorene and squaraine monomers were developed. These Pdotsthat emit in the NIR region and can be used, e.g., for biologicalapplications. FIG. 24D, for example, shows a schematic illustration of asquaraine based narrow emissive polymer dots and Pdot-bioconjugates forspecific cellular targeting. With their nanometer-scale particle sizes,high quantum yields in water, and small FWHM (<40 nm), these Pdots can,e.g., be used for multiplexing the simultaneous targeting of severalmolecules on the surfaces of cells. FIG. 25E shows photophysical dataand particle size of the narrow-band emissive PFS Pdots and PFS 5.5Pdots with varying squaraine ratio. FIG. 25F shows the fluorescenceemission spectra of the narrow-band emissive PFS Pdots and PFS 5.5 Pdotswith varying squaraine ratio. The PFS-1.5% squaraine copolymer Pdotsshows emission FWHM of 37 nm at 690 nm and fluorescence quantum yield is0.30. FIG. 25G shows absorption and fluorescence spectra of PFS andPFS5.5 Pdots with 1.5% squaraine molar ratio. Top-left panel a shows theabsorption and fluorescence spectra of PFS Pdot with 1.5% squarainemolar ratio. Top-right panel b shows the fluorescence spectra of PFSPdot at 1.5% molar ratio of squaraine dye in THF and the Pdots formed inwater. Bottom-left panel c shows the absorption spectra of PFS and PFS5.5 Pdots (with 1.5% squaraine molar ratio) in water. Bottom-right paneld shows the fluorescence spectra of PFS and PFS 5.5 Pdots (with 1.5%squaraine molar ratio) in water. FIG. 25H shows the particle sizedistributions of PFS Pdots (average size of 19 nm) and PFS 5.5 Pdots(average size of 19 nm). Bottom panels show the TEM images of PFS Pdots(c) and PFS 5.5 Pdots (d). FIG. 25I shows single-particle brightnessimages with 405 nm excitation for three samples: (a) Qdots 705; (b) 1.5%PFS Pdots; (c) 1.5% PFS 5.5 Pdots. The images in the top panel wereobtained under identical excitation and detection conditions. All scalebars represent 5 μm. The bottom panels show the histograms of thebrightness distributions. FIG. 25J shows flow cytometry intensitydistributions of MCF-7 breast cancer cells labeled with Qdot705-streptavidin, PFS Pdot-streptavidin, and PFS5.5 Pdot-streptavidin.FIG. 25K shows confocal fluorescence images of MCF-7 cells labeled withPFS Pdot-streptavidin and PFS 5.5 Pdot-streptavidin probes.

Example 9 Narrow-band Emissive Pdots Embedded with Quantum Dot (QD)

The present example provides a method for obtaining a narrow-bandemissive Pdots embedded with inorganic quantum dots (QDs).

As shown in FIG. 26A, we synthesized a general semiconducting polymerPFBT functionalized with amino groups and then converted these aminogroups to thiols by 2-iminothiolane (Traut's reagent) under THF in aneffort to have an efficient ligand-exchange process with original aminocapping ligands on QD surfaces. After the exchange reaction, we blendeda copolymer, PS-PEG-COOH that consists of carboxyl groups for furtherbiological applications, and then co-precipitated polymer-QD mixtures inwater under sonication to from Pdot-QD NPs. From transmission electronmicroscopy (TEM) images (FIG. 1B), we can clearly see that a cluster ofQDs was encapsulated by polymers in which there are ˜30 QDs embeddedinside each individual Pdot-Qdot nanoparticle. We found that thecrystalline structure of QDs was preserved intact during the preparationprocesses (FIG. 26B) and remains optically stable and biologicallyactive even after two months of storage at 4° C. in physiological pHbuffers. Dynamic light scattering (DLS) measurements also showed thatthe average diameter of Pdot-Qdot NPs was 25 nm (FIG. 26C). We noticedthat the color of emission changed drastically from bright yellowfluorescence before nanoprecipitation to deep red emission afternanoprecipitation. The yellow fluorescence is characteristic of PFBT,while the red fluorescence is from QDs under UV light irradiation. Thisphenomenon is indicative of a highly efficient energy from PFBT to QDsonly when they are in close proximity.

FIG. 27A shows the absorption spectrum of Pdot-Qdot NPs. The absorptionpeak at ˜450 nm (blue arrow) is evidence of PFBT and the smallabsorption at ˜640 nm (red arrow) is from CdSe QDs, while the highabsorption below 400 nm is attributed to both PFBT and QDs. In thepresent work, we employed three different sizes of QDs that emit at 655nm (QD655), 705 nm (QD705), and 800 nm (QD800), respectively. Wecarefully optimized the quantitative ratio of PFBT to QD in order tohave an efficient energy transfer from PFBT to QDs while keeping theircompact size (i.e. minimal number of QDs inside a Pdot). As shown inFIG. 27B, the fluorescence signal of PFBT was almost completelyquenched, indicating an efficient energy/electron transfer from PFBT toQDs. More importantly, the emission bandwidth of Pdot-Qdot NPs remainedunaltered, allowing us to create Pdot-based NPs with narrow emission.Taking QD655 for example, the full width at half maximum of Pdot-QD655in water is ˜25 nm (solid red line in FIG. 27B), which is almost thesame as the original QD655 in decane (dashed red line). Moreover, wehave demonstrated this technique can be applied to NIR QDs such as QD705and QD800 (purple and pink lines in FIG. 27B). The quantum yields ofPdot-QD655, Pdot-QD705, and Pdot-QD800 in buffer solutions weredetermined to be 23%, 38%, and 29%, respectively. This implies that weshould be able to take the advantage of the unique optical properties ofQDs such as narrow-band and NIR emission, while implanting the merits ofPdots on them, including the large absorption cross-sections and facilesurface functionalization. To prove our concept, we first performed theexperiments of single particle fluorescence brightness. We have shownthat PFBT-DBT Pdots emitting at ˜650 nm are 15 times brighter thansingle QD655. Here, we directly compared the single-particle brightnessof PFBT-DBT Pdots with PFBT-QD655 NPs. We found the particle brightnessof PFBT-QD655 is comparable to that of PFBT-DBT (FIG. 27C, 27D). Becausethe particle brightness is given by the product of absorption crosssection and quantum yield and in this case the quantum yield was onlyslightly altered, we can then attribute the brightness enhancement tothe vast increase in optical absorption cross section originated frompolymer coating or/and QD multiples. To evaluate the bioconjugationactivity of this hybrid material, we performed the subcellularmicrotubule labelling in HeLa cells. We first bioconjugate streptavidinonto Pdot-Qdot NP surfaces via1-ethyl-3-[3-dimethylaminopropyl]carbodiimidehydrochloride-(EDC)-catalyzed coupling).

FIG. 28 shows the two-color confocal microscopy images of microtubulesin HeLa cells labeled with Pdot-QD705-streptavidin. The bluefluorescence is from nuclear counter-stain Hoechst 34580 (A), the redfluorescence (B) is from Pdot-QD705-streptavidin, and (C) is the overlayof panels (A) and (B). (D-F) are images of the control sample wherecells were incubated with Pdot-QD705-streptavidin but in the absence ofbiotinylated primary antibody. As indicated by the fluorescence images,the Pdot-QD-streptavidin conjugates can specifically label the cellulartargets. FIG. 28G-H show the flow cytometry results of Pdot-Qdot labeledMCF-7 cells. The purple and pink lines show the fluorescence intensitydistributions of Pdot-QD705-streptavidin and Pdot-QD800-streptavidinlabeled cells, respectively. The black lines represent the results ofcontrol samples (no primary Biotin anti-human CD326 EpCAM antibody).Again, these results show specific labeling of the Pdot-QD probeswithout nonspecific labeling.

Example 10 Narrow-band Emission from Blended Pdots Comprising aNarrow-band Emissive Fluorene-BODIPY Copolymer

The present example provides a method for obtaining narrow-band emissionby using blended Pdots comprising a narrow-band emissive fluorene-BODIPYcopolymer.

A narrow-band emissive copolymer was first synthesized by using theBODIPY monomer 2a in FIG. 12 as narrow-band monomer and fluorene as ageneral monomer. This copolymer in a good solvent such as THF exhibitnarrow-band emission. However, the Pdots prepared by this polymerexhibit broad-band emission (FIG. 29B) because the fluorophores weredensely packed and aggregation states were formed. We used a blendingstrategy to obtain narrow-band emission. The narrow-band emissiveFluorene-BODIPY polymer was mixed with a conventional broad-bandemission semiconducting polymer PFBT in THF, and blended Pdots wereprepared by injecting a THF solution of the polymer mixture into water.As indicated by the emission spectrum (FIG. 29C), the blended Pdotsexhibit narrow-band emission as compared to the pure Fluoene-BODIPYPdots. The narrow-band emissive Fluoene-BODIPY polymer was dispersed inthe PFBT host, therefore preventing the formation of aggregation. Thereis also efficient intra-particle energy transfer from PFBT to the BODIPYunits. As a result, the PFBT emission was completely quenched, and theblended Pdots gave narrow-band emission.

It is also possible to use other fluorescent polymers, copolymers, andin particular other BODIPY fluorescent copolymers such as the ones shownin FIG. 29D. FIG. 29F-29H corresponds to the fluorescence spectra ofchromophoric polymer dots formed using the different copolymers andpolymer blends shown in FIG. 29D (I-IV). It is evident from this examplethat different types of copolymers and blends of polymers and copolymerscan be used, and also that blending can significantly improve thequantum yield of the resultant chromophoric polymer dots. Thesechromophoric polymer dots were formed from the polymers shown in FIG.29D, where a solution of polymer precursor mixture in THF (for example,1.2 mL, 100 ppm of PFBT solution, 0.6 mL, 100 pm of PF-5TBT solution,0.2 mL, 100 ppm of 10% mol deep red BODIPY fluorene copolymer solution,0.5 mL, 100 ppm of PSMA1800) was quickly injected into water (10 mL)under ultrasonication. THF was evaporated by N₂ flow at 60° C. and thesolution was concentrated to 8-9 mL.

Example 11 Narrow-band Emission Chromophoric Polymer Dots Formed fromPFPPyBPh

FIG. 30A shows the synthetic scheme ofPoly[(9,9-dioctylfluoren-2,7-yl)-alt-(diphenyl-2-(2-pyridinyl-κN)phenyl-κC]-(T-4)-boron-5,5-yl)](PFPPyBPh). Briefly, Compound 2: To a solution of phenyl lithium (1.7mL, 1.8 M in dibutyl ether) was added ZnCl₂ (3.3 mL, 1 M in diethylether) at 0° C. The solution was stirred for 30 min at 0° C., and for 1hour at room temperature. The compound 1 (0.6 g) and toluene (30 mL)were added in one portion. Then, the resultant solution was stirred at80° C. overnight. After cooling to room temperature, the solution waspoured into water. The organic phase was separated and the aqueous phasewas extracted with dichloromethane twice. The combined organic phaseswere dried over anhydrous Na₂SO₄. After removing the solvent, the crudeproduct was purified by silica column to give a white solid (0.3 g,50%). ¹H NMR (CDCl₃, ppm): 8.57 (dd, 1H), 8.20 (dd, 1H), 7.93 (dd, 1H),7.82 (d, 1H), 7.74 (d, 1H), 7.50 (dd, 1H), 7.26-7.18 (m, 10H).

Polymer PFPPyBPh: In a 50-mL single-neck flask, the compounds 3 and 2were charged. Toluene (3.5 mL), aqueous Na₂CO₃ (2 mL, 2M) and A336 (2drops) were added consequently. The solution was degassed twice beforeadding Pd(PPh₃)₄ (7.5 mg). Then, the solution was heated to 120° C. for48 hours. After cooling to room temperature, the solution was pouredinto methanol. The resultant solid was dissolved into chloroform andpassed through a short column. The concentrated solution was then pouredinto methanol. The solid was collected by filtration and dried undervacuum overnight (120 mg, 68%).

FIG. 30B shows the absorption and fluorescence spectra of thesynthesized PFPPyBPh polymer in THF (tetrahydrofuran). FIG. 30C showsthe absorption and fluorescence spectra of the chromophoric polymer dotsformed using the synthesized PFPPyBPh polymer. The formed chromophoricpolymer dots, which also contained some PSMA, were measured by dynamiclight scattering to be around 15 nm in diameter.

Example 12 Synthesis of BODIPY Monomer 4a (Shown in FIG. 12) and RelatedNarrow-band Emissive Polymer with Side-chain Amine Groups

The present example provides a method for obtaining narrow-band BODIPYmonomer4a and a narrow-band emissive fluorene-BODIPY copolymerpolymer540 with side-chain amine groups.

Synthesis of BODIPY monomer 4a. To a solution of4-methyl-3,5-diiodobenzaldehyde (1.5 g, 4.2 mmol) and2,4-dimethyl-3-ethyl-1H-pyrrole (1 g, 10.5 mmol) in dry CH₂Cl₂ (120 ml)was added a solution of 110 μl trifluoroacetic acid in dry CH₂Cl₂ (5 ml)slowly at room temperature. 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone(0.95 g, 4.2 mmol) was added after 3 h stirring under ice bath coolingand stirred for 10 min. The solution was stirred for an additional 1 hat room temperature. NEt₃ (10 ml, 72 mmol) was added, followed by slowaddition of BF₃. Et₂O (12 ml, 81 mmol). The reaction mixture was washedafter 10 h of stirring at room temperature with saturated aqueous Na₂CO₃solution (2×100 ml), dried over Na₂SO₄, and concentrated on a rotaryevaporator. The brown, oily residue was purified by columnchromatography on silica with hexane/CH₂Cl₂=3:1. The product fractionwith greenish fluorescence was dried to yield an orange solid. Yield:0.48 g, 19.5%. ¹H NMR (CDCl₃, 500MHz): δ=7.831 (s, 2H), 6.042 (s, 2H),2.874 (s, 3H), 2.581 (s, 6H), 1.544 (s, 6H). ¹³C NMR (CDCl₃, 125 MHz):δ=156.25, 144.12, 142.83, 138.94, 135.89, 131.11, 121.67, 99.09, 34.93,15.14, 14.61. HRMS (ESI) (M⁺, C₂₄H₂₇BF₂I₂N₂): calcd, 647.0442; found,647.0432.

Synthesis of fluorene-BODIPY polymer540 with side-chain amine groups(PF5%540BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 372mg (1.35 mmol) of bis(1,5-cyclooctadiene) nickel(0), 150 mg (1.35 mmol)of cyclooctadiene, and 210 mg (1.35 mmol) of bypyridine in 5.0 mL of a1:1 mixture of toluene and dimethylformamide (DMF). A dark purple colorthen developed. The solution was heated to 60° C. In the glovebox, a dry20 mL flask was charged with 19.4 mg (0.03 mmol) of BODIPY monomer 1a,299.4 mg (0.546 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and 15.3 mg(0.024 mmol) of monomer 4a in 4.0 mL of a 1:1 mixture of toluene andDMF, then they were added dropwise into the above catalyst mixture. Theflask containing this solution was covered with foil to protect it fromlight and the reaction mixture was refluxed for 4 days. Then 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 12 h at 60° C. The product was diluted with 50ml of toluene and washed with aqueous 15 wt % of sodium thiosulfatesolution (3×50 mL) followed by washing with Milli-Q water and dryingover MgSO₄, for the removal of residual iodine from polymer. The polymersolution was evaporated and dissolved in dichloromethane. After polymersolution was filtered, the concentrated polymer solution indichloromethane was poured into 100 ml of MeOH, and filtered. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24h, and filtered. Polymer was obtained as orange solid (120 mg).De-protection of amine group from the polymer was finished as thefollowing procedure, polymer was dissolved in DCM (50 mL) and TFA (1.5mL) was added to remove protecting groups and generate amine groups. Themixture was stirred at room temperature overnight (in dark) and thenwashed with 10% NaOH water solution three times. DCM phase wasseparated, concentrated to ca. 5 mL, and then added into methanol (80mL) to precipitate the final polymer. Yield: 130 mg, 54.2%. ¹H NMR(CDCl₃, 500 MHz): δ=7.90-7.75 (m), 7.53 (m), 7.42-7.43 (m, 6H), 6.08 (m,2H), 2.64 (s, 6H), 2.18 (s, 4H), 1.63 (s, 6H), 1.21 (s, 24H), 0.88 (s,6H). ¹³C NMR (CDCl₃, 125 MHz): δ=151.87, 140.57, 140.07, 126.21, 121,53,120.02, 55.40, 40.46, 31.86, 30.1, 29.78, 29.3, 23.99, 22.66, 14.15. Mn:57512, Mw: 90491, PDI: 1.573.

Example 13 Synthesis of Two General Fluorescent Polymers with Side-ChainAmine Groups for Chemically Cross-Linking with Narrow-band EmissivePolymer

The present example provides a method for obtaining general fluorescentpolymer with side-chain amine groups that can chemically cross-link withnarrow-band emissive polymers.

Synthesis of PF10BT with amine group (PF10BT4NH₂). Polymer issynthesized by palladium-catalyzed Suzuki coupling reaction from9,9-dioctylfluorene and 4,7-Dibromobenzo[c]-1,2,5-thiadiazole. 197.4 mg(0.36 mmol) of 9,9-Dioctyl-2,7-dibromofluorene, 279.2 mg (0.5 mmol) of9,9-dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester, 29.4mg (0.1 mmol) of 4,7-dibromobenzo[c]-1,2,5-thiadiazole, 25.5 mg (0.04mmol) of 2,7-dibromo-9,9-bis(3-(tert-butyl hexylcarbamate)fluorine, 2drops of aliquot 336, 10 ml of 2M Na₂CO₃ aqueous solution, 15 ml oftoluene were placed in a 50 ml flask. The flask was evacuated andrefilled with N₂ four times by using the freeze/thaw method andPd(PPh₃)₄ (1-1.5 mol %) was added. The flask was further degassed fourtimes, then reaction was heated to 100° C. and stirred under N₂. After70 h 0.2 ml of bromobenzene and 15 mg of phenylboronic acid in toluenewere added to end-cap the polymer chain and the reaction was stirred foran additional 2 h at 100° C. respectively. The whole mixture was pouredinto 100 ml of MeOH, filtered, and washed with 0.2M of HCl. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24 hand dried in vacuum oven to obtain dark yellow solid. De-protection ofamine group from the polymer was finished as the following procedure,polymer was dissolved in DCM (50 mL) and TFA (1.5 mL) was added toremove protecting groups and generate amine groups. The mixture wasstirred at room temperature overnight (in dark) and then washed with 10%NaOH water solution three times. DCM phase was separated, concentratedto ca. 10 mL, and then added into methanol (80 mL) to precipitate thefinal polymer. Yield: 320 mg, 79%.

Synthesis of PF5TBT with amine group (PF5TBT4NH₂). Polymer issynthesized by palladium-catalyzed Suzuki coupling reaction from9,9-dioctylfluorene and4,7-Bis(2-bromo-5-thienyl)-2,1,3-benzothiadiazole. 224.8 mg (0.41 mmol)of 9,9-Dioctyl-2,7-dibromofluorene, 279.2 mg (0.5 mmol) of9,9-dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester, 22.9mg (0.05 mmol) of 4,7-bis(2-bromo-5-thienyl)-2,1,3-benzothiadiazole,25.5 mg (0.04 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorine, 2 drops of aliquot 336, 10 ml of 2M Na₂CO₃aqueous solution, 15 ml of toluene were placed in a 50 ml flask. Theflask was evacuated and refilled with N₂ four times by using thefreeze/thaw method and Pd(PPh₃)₄ (1-1.5 mol %) was added. The flask wasfurther degassed four times, then reaction was heated to 100° C. andstirred under N₂. After 70 h 0.2 ml of bromobenzene and 15 mg ofphenylboronic acid in toluene were added to end-cap the polymer chainand the reaction was stirred for an additional 2 h at 100° C.respectively. The whole mixture was poured into 100 ml of MeOH,filtered, and washed with 0.2M of HCl. The precipitate was stirred in 50ml of acetone at room temperature for 24 h and dried in vacuum oven toobtain dark yellow solid. De-protection of amine group from the polymerwas finished as the following procedure, polymer was dissolved in DCM(50 mL) and TFA (1.5 mL) was added to remove protecting groups andgenerate amine groups. The mixture was stirred at room temperatureovernight (in dark) and then washed with 10% NaOH water solution threetimes. DCM phase was separated, concentrated to ca. 10 mL, and thenadded into methanol (80 mL) to precipitate the final polymer. Yield: 297mg, 71%.

Example 14 Synthesis of BODIPY Monomer 5a (Shown in FIG. 12) and RelatedNarrow-band Emissive Polymer with Side-chain Amine Groups

The present example provides a method for obtaining narrow-band BODIPYmonomer5a and a narrow-band emissive fluorene-BODIPY copolymerpolymer570 with side-chain amine groups.

Synthesis of BODIPY monomer 5a. To a solution of4-methyl-3,5-diiodobenzaldehyde (0.69 g, 1.9 mmol) and2,3-Tetramethylenepyrrole (0.5 g, 4.1 mmol) in dry CH₂Cl₂ (100 ml) wasadded a solution of 90 μl trifluoroacetic acid in dry CH₂Cl₂ (5 ml)slowly at room temperature. 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone(0.42 g, 2.0 mmol) was added after 3 h stirring under ice bath coolingand stirred for 10 min. The solution was stirred for an additional 1 hat room temperature. NEt₃ (5 ml, 36 mmol) was added, followed by slowaddition of BF₃. Et₂O (6 ml, 40 mmol). The reaction mixture was washedafter 10 h of stirring at room temperature with saturated aqueous Na₂CO₃solution (2×50 ml), dried over Na₂SO₄, and concentrated on a rotaryevaporator. The brown, oily residue was purified by columnchromatography on silica with hexane/CH₂Cl₂=3:1. The product fractionwith greenish fluorescence was dried to yield a red solid. Yield: 146mg, 12%.

Synthesis of Fluorene-BODIPY polymer570 with an amine group(PF5%570BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 316mg (1.15 mmol) of bis(1,5-cyclooctadiene) nickel(0), 128 mg (1.15 mmol)of cyclooctadiene, and 178 mg (1.15 mmol) of bypyridine in 5.0 mL of a1:1 mixture of toluene and dimethylformamide (DMF). A dark purple colorthen developed. The solution was heated to 60° C. In the glovebox, a dry20 mL flask was charged with 32.2 mg (0.025 mmol) of BODIPY monomer 5a,250 mg (0.455 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and 12.8 mg (0.02mmol) of 2,7-dibromo-9,9-bis(3-(tert-butyl hexylcarbamate)fluorine in4.0 mL of a 1:1 mixture of toluene and DMF, then they were addeddropwise into the above catalyst mixture. The flask containing thissolution was covered with foil to protect it from light and the reactionmixture was refluxed for 4 days. Then 4 drops of iodobenzene was addedto end-cap the polymer chain and the reaction was stirred for anadditional 12 h at 60° C. The product was diluted with 50 ml of tolueneand washed with aqueous 15 wt % of sodium thiosulfate solution (3×50 mL)followed by washing with Milli-Q water and drying over MgSO₄, for theremoval of residual iodine from polymer. The polymer solution wasevaporated and dissolved in dichloromethane. After polymer solution wasfiltered, the concentrated polymer solution in dichloromethane waspoured into 100 ml of MeOH, and filtered. The precipitate was stirred in50 ml of acetone at room temperature for 24 h, and filtered. Polymer wasobtained as red solid. De-protection of amine group from the polymer wasfinished as the following procedure, polymer was dissolved in DCM (50mL) and TFA (1.5 mL) was added to remove protecting groups and generateamine groups. The mixture was stirred at room temperature overnight (indark) and then washed with 10% NaOH water solution three times. DCMphase was separated, concentrated to ca. 5 mL, and then added intomethanol (80 mL) to precipitate the final polymer. Yield: 116 mg, 58%.

Example 15 Synthesis of BODIPY Monomer 6a (Shown in FIG. 12) and RelatedNarrow-band Emissive Polymer with Side-chain Amine Groups

The present example provides a method for obtaining narrow-band BODIPYmonomer5a and a narrow-band emissive fluorene-BODIPY copolymerpolymer600 with side-chain amine groups.

Synthesis of BODIPY monomer 6a. p-tolualdehyde (120 mg, 0.96 mmol),BODIPY monomer 4a (400 mg, 0.62 mmol), p-toluene sulfonic acid (60 mg),2 ml of acetic acid, and piperidine (3 ml) were dissolved in 30 ml ofbenzene refluxed for 10 h by using a Dean-Stark apparatus. The mixturewas cooled to room temperature, the solvents were removed under vacuum,and the crude product was purified by column chromatography on silicagel eluted with ethyl acetate/hexane 1:7. The crude was recrystallizedfrom chloroform/methanol to give the product as a metallic shiny solid.Yield: 70 mg, 15%.

Synthesis of fluorene-BODIPY polymer600 with amine group(PF5%600BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 248mg (0.9 mmol) of bis(1,5-cyclooctadiene) nickel(0), 97 mg (0.9 mmol) ofcyclooctadiene, and 140 mg (0.9 mmol) of bypyridine in 4.0 mL of a 1:1mixture of toluene and dimethylformamide (DMF). A dark purple color thendeveloped. The solution was heated to 60° C. In the glovebox, a dry 20mL flask was charged with 15.2 mg (0.02 mmol) of BODIPY monomer 6a,199.6 mg (0.364 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and 10.2 mg(0.016 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorine in 4.0 mL of a 1:1 mixture of toluene and DMF,then they were added dropwise into the above catalyst mixture. The flaskcontaining this solution was covered with foil to protect it from lightand the reaction mixture was refluxed for 4 days. Then 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 12 h at 60° C. The product was diluted with 50ml of toluene and washed with aqueous 15 wt % of sodium thiosulfatesolution (3×50 mL) followed by washing with Milli-Q water and dryingover MgSO₄, for the removal of residual iodine from polymer. The polymersolution was evaporated and dissolved in dichloromethane. After polymersolution was filtered, the concentrated polymer solution indichloromethane was poured into 100 ml of MeOH, and filtered. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24h, and filtered. Polymer was obtained as red solid. De-protection ofamine group from the polymer was finished as the following procedure,polymer was dissolved in DCM (50 mL) and TFA (1.0 mL) was added toremove protecting groups and generate amine groups. The mixture wasstirred at room temperature overnight (in dark) and then washed with 10%NaOH water solution three times. DCM phase was separated, concentratedto ca. 5 mL, and then added into methanol (80 mL) to precipitate thefinal polymer. Yield: 105 mg, 65.6%.

Example 16 Synthesis of a Fluorescent Polymers with Side-chain AmineGroups for Chemically Cross-linking with Narrow-band Emissive Polymer

The present example provides a method for obtaining a fluorescentpolymer with side-chain amine groups that can chemically cross-link withnarrow-band emissive polymers.

Synthesis of PF47BSeD with amine group (PF47BSeD3NH₂). Polymer issynthesized by palladium-catalyzed Suzuki coupling reaction from9,9-dioctylfluorene and 4,7-dibromobenzo[c]-1,2,5-selenadiazole. 224.8mg (0.41 mmol) of 9,9-dioctyl-2,7-dibromofluorene, 139.6 mg (0.25 mmol)of 9,9-dioctylfluorene-2,7-diboronic acid bis(1,3-propanediol) ester,80.1 mg (0.235 mmol) of 4,7-dibromobenzo[c]-1,2,5-selenadiazole, 9.57 mg(0.015 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorene, 2 drops of aliquot 336, 10 ml of 2M Na₂CO₃aqueous solution, 15 ml of toluene were placed in a 50 ml flask. Theflask was evacuated and refilled with N₂ four times by using thefreeze/thaw method and Pd(PPh₃)₄ (1-1.5 mol %) was added. The flask wasfurther degassed four times, then reaction was heated to 100° C. andstirred under N₂. After 70 h 0.2 ml of bromobenzene and 15 mg ofphenylboronic acid in toluene were added to end-cap the polymer chainand the reaction was stirred for an additional 2 h at 100° C.respectively. The whole mixture was poured into 100 ml of MeOH,filtered, and washed with 0.2M of HCl. The precipitate was stirred in 50ml of acetone at room temperature for 24 h and dried in vacuum oven toobtain dark yellow solid. De-protection of amine group from the polymerwas finished as the following procedure, polymer was dissolved in DCM(50 mL) and TFA (1.0 mL) was added to remove protecting groups andgenerate amine groups. The mixture was stirred at room temperatureovernight (in dark) and then washed with 10% NaOH water solution threetimes. DCM phase was separated, concentrated to ca. 10 mL, and thenadded into methanol (80 mL) to precipitate the final polymer. Yield: 114mg, 76%.

Example 17 Synthesis of BODIPY Monomer 7a (Shown in FIG. 12) and RelatedNarrow-band Emissive Polymer with Side-chain Amine Groups

The present example provides a method for obtaining narrow-band BODIPYmonomer7a and a narrow-band emissive fluorene-BODIPY copolymerpolymer655 with side-chain amine groups.

Synthesis of BODIPY monomer 5a. p-tolualdehyde (168 mg, 1.4 mmol),BODIPY monomer 1a (200 mg, 0.34 mmol), p-toluene sulfonic acid (50 mg),3 ml of acetic acid, and piperidine (3 ml) were dissolved in 30 ml ofbenzene refluxed for 10 h by using a Dean-Stark apparatus. The mixturewas cooled to room temperature, the solvents were removed under vacuum,and the crude product was purified by column chromatography on silicagel eluted with ethyl acetate/hexane 1:7. The crude was recrystallizedfrom chloroform/methanol to give the product as a metallic shiny solid.Yield: 99 mg, 37%.

Synthesis of fluorene-BODIPY polymer655 with amine group(PF5%655BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 248mg (0.9 mmol) of bis(1,5-cyclooctadiene) nickel(0), 97 mg (0.9 mmol) ofcyclooctadiene, and 140 mg (0.9 mmol) of bypyridine in 4.0 mL of a 1:1mixture of toluene and dimethylformamide (DMF). A dark purple color thendeveloped. The solution was heated to 60° C. In the glovebox, a dry 20mL flask was charged with 15.9 mg (0.02 mmol) of BODIPY monomer 7a,199.6 mg (0.364 mmol) of 9,9-dioctyl-2,7-dibromofluorene and 10.2 mg(0.016 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorine in 4.0 mL of a 1:1 mixture of toluene and DMF,then they were added dropwise into the above catalyst mixture. The flaskcontaining this solution was covered with foil to protect it from lightand the reaction mixture was refluxed for 4 days. Then 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 12 h at 60° C. The product was diluted with 50ml of toluene and washed with aqueous 15 wt % of sodium thiosulfatesolution (3×50 mL) followed by washing with Milli-Q water and dryingover MgSO₄, for the removal of residual iodine from polymer. The polymersolution was evaporated and dissolved in dichloromethane. After polymersolution was filtered, the concentrated polymer solution indichloromethane was poured into 100 ml of MeOH, and filtered. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24h, and filtered. Polymer was obtained as red solid. De-protection ofamine group from the polymer was finished as the following procedure,polymer was dissolved in DCM (50 mL) and TFA (1.0 mL) was added toremove protecting groups and generate amine groups. The mixture wasstirred at room temperature overnight (in dark) and then washed with 10%NaOH water solution three times. DCM phase was separated, concentratedto ca. 5 mL, and then added into methanol (80 mL) to precipitate thefinal polymer. Yield: 97 mg, 60.6%.

Example 18 Synthesis of BODIPY Monomer 8a (Shown in FIG. 12) and RelatedNarrow-band Emissive Polymer with Side-chain Amine Groups

The present example provides a method for obtaining narrow-band BODIPYmonomer8a and a narrow-band emissive fluorene-BODIPY copolymer polymerswith side-chain amine groups.

Synthesis of BODIPY monomer 8a. p-Tolualdehyde (300 mg, 2.5 mmol),BODIPY monomer 4a (400 mg, 0.62 mmol), p-toluene sulfonic acid (80 mg),2 ml of acetic acid, and piperidine (3 ml) were dissolved in 35 ml ofbenzene refluxed for 10 h by using a Dean-Stark apparatus. The mixturewas cooled to room temperature, the solvents were removed under vacuum,and the crude product was purified by column chromatography on silicagel eluted with ethyl acetate/hexane 1:7. The crude was recrystallizedfrom chloroform/methanol to give the product as a metallic shiny solid.Yield: 180 mg, 33%.

Synthesis of fluorene-BODIPY polymer670 with amine group(PF5%670BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 248mg (0.9 mmol) of bis(1,5-cyclooctadiene) nickel(0), 97 mg (0.9 mmol) ofcyclooctadiene, and 140 mg (0.9 mmol) of bypyridine in 4.0 mL of a 1:1mixture of toluene and dimethylformamide (DMF). A dark purple color thendeveloped. The solution was heated to 60° C. In the glovebox, a dry 20mL flask was charged with 17.6 mg (0.02 mmol) of BODIPY monomer 8a,199.6 mg (0.364 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and 10.2 mg(0.016 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorine in 4.0 mL of a 1:1 mixture of toluene and DMF,then they were added dropwise into the above catalyst mixture. The flaskcontaining this solution was covered with foil to protect it from lightand the reaction mixture was refluxed for 4 days. Then 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 12 h at 60° C. The product was diluted with 50ml of toluene and washed with aqueous 15 wt % of sodium thiosulfatesolution (3×50 mL) followed by washing with Milli-Q water and dryingover MgSO₄, for the removal of residual iodine from polymer. The polymersolution was evaporated and dissolved in dichloromethane. After polymersolution was filtered, the concentrated polymer solution indichloromethane was poured into 100 ml of MeOH, and filtered. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24h, and filtered. Polymer was obtained as red solid. De-protection ofamine group from the polymer was finished as the following procedure,polymer was dissolved in DCM (50 mL) and TFA (1.0 mL) was added toremove protecting groups and generate amine groups. The mixture wasstirred at room temperature overnight (in dark) and then washed with 10%NaOH water solution three times. DCM phase was separated, concentratedto ca. 5 mL, and then added into methanol (80 mL) to precipitate thefinal polymer. Yield: 109 mg, 68.1%.

Synthesis of fluorene-BODIPY polymer680 with amine group(PF5%680BODIPY4NH₂). In a glovebox under nitrogen atmosphere, a drythree neck 50 mL round-bottom flask with stir bar was charged with 248mg (0.9 mmol) of bis(1,5-cyclooctadiene) nickel(0), 97 mg (0.9 mmol) ofcyclooctadiene, and 140 mg (0.9 mmol) of bypyridine in 4.0 mL of a 1:1mixture of toluene and dimethylformamide (DMF). A dark purple color thendeveloped. The solution was heated to 60° C. In the glovebox, a dry 20mL flask was charged with 16.5 mg (0.02 mmol) of BODIPY monomer 3a,199.6 mg (0.364 mmol) of 9,9-Dioctyl-2,7-dibromofluorene and 10.2 mg(0.016 mmol) of 2,7-dibromo-9,9-bis(3-(tert-butylhexylcarbamate)fluorine in 4.0 mL of a 1:1 mixture of toluene and DMF,then they were added dropwise into the above catalyst mixture. The flaskcontaining this solution was covered with foil to protect it from lightand the reaction mixture was refluxed for 4 days. Then 4 drops ofiodobenzene was added to end-cap the polymer chain and the reaction wasstirred for an additional 12 h at 60° C. The product was diluted with 50ml of toluene and washed with aqueous 15 wt % of sodium thiosulfatesolution (3×50 mL) followed by washing with Milli-Q water and dryingover MgSO₄, for the removal of residual iodine from polymer. The polymersolution was evaporated and dissolved in dichloromethane. After polymersolution was filtered, the concentrated polymer solution indichloromethane was poured into 100 ml of MeOH, and filtered. Theprecipitate was stirred in 50 ml of acetone at room temperature for 24h, and filtered. Polymer was obtained as red solid. De-protection ofamine group from the polymer was finished as the following procedure,polymer was dissolved in DCM (50 mL) and TFA (1.0 mL) was added toremove protecting groups and generate amine groups. The mixture wasstirred at room temperature overnight (in dark) and then washed with 10%NaOH water solution three times. DCM phase was separated, concentratedto ca. 5 mL, and then added into methanol (80 mL) to precipitate thefinal polymer. Yield: 122 mg, 76.2%.

Example 19 Formation of Narrow-band Emissive Polymer Dots by ChemicallyCross-linking General Fluorescent Polymer with Narrow-band EmissivePolymer

The present example provides a method for obtaining narrow-band emissivepolymer dots by chemically cross-linking general fluorescent polymerwith narrow-band fluorene-BODIPY polymers. The synthesis scheme is shownin FIG. 31.

Cross-linking reaction of PF5%540BODIPY4NH₂ and PF10BT4NH₂ with PSMA. 1mg of PF5%540BODIPY4NH₂ and 1 mg of PF10BT-4NH₂ were dissolved in 2 mLof THF, then 125 μL (4000 ppm) of 0.5 PSMA-8000 (33% MA, 67% PS, Mw:8000) in THF was added into the above THF solution. The solution wascovered with foil to protect it from light and stirred for 72 h at roomtemperature. 100-200 μL of above solution was diluted to 40-50 ppm withTHF and was used for Pdots preparation. FIG. 42A shows the absorptionand fluorescence spectra of resulting crosslinked Pdots. The crosslinked540 Pdots emission exhibit a FWHM of 39 nm.

Crosslinking reaction of PF5%570BODIPY4NH₂ and PF46BT4NH₂ with PSMA. 0.7mg of PF5%570BODIPY4NH₂ and 1.3 mg of PF10BT-4NH₂ were dissolved in 2 mLof THF, then 125 μL (4000 ppm) of 0.5 PSMA-8000 (33% MA, 67% PS, Mw:8000) in THF was added into the above THF solution. The solution wascovered with foil to protect it from light and stirred for 72 h at roomtemperature. 100-200 μL of above solution was diluted to 40-50 ppm withTHF and was used for Pdots preparation. FIG. 42B shows the absorptionand fluorescence spectra of resulting crosslinked Pdots. The crosslinked570 Pdots emission exhibit a FWHM of 37 nm.

Cross-linking reaction of PF5%590BODIPY4NH₂ and PF46BT4NH₂ with PSMA.0.8 mg of PF5%570BODIPY4NH₂ and 1.2 mg of PF46BT-4NH₂ were dissolved in2 mL of THF, then 125 μL (4000 ppm) of 0.5 PSMA-8000 (33% MA, 67% PS,Mw: 8000) in THF was added into the above THF solution. The solution wascovered with foil to protect it from light and stirred for 72 h at roomtemperature. 100-200 μL of above solution was diluted to 40-50 ppm withTHF and was used for Pdots preparation. FIG. 42A shows the absorptionand fluorescence spectra of resulting crosslinked Pdots. The crosslinked590 Pdots emission exhibit a FWHM of 55 nm.

Cross-linking reaction of PF5%600BODIPY4NH₂ and PF47BSeD4NH₂ with PSMA.0.8 mg of PF5%600BODIPY4NH₂ and 1.2 mg of PF46BT-4NH₂ were dissolved in2 mL of THF, then 125 μL (4000 ppm) of 0.5 PSMA-8000 (33% MA, 67% PS,Mw: 8000) in THF was added into the above THF solution. The solution wascovered with foil to protect it from light and stirred for 72 h at roomtemperature. 100-200 μL of above solution was diluted to 40-50 ppm withTHF and was used for Pdots preparation. FIG. 42C shows the absorptionand fluorescence spectra of resulting crosslinked Pdots. The crosslinked600 Pdots emission exhibit a FWHM of 38 nm.

Cross-linking reaction of PF5%655BODIPY4NH₂, PF46BT4NH₂ and PF5TBT4NH₂with PSMA. 0.4 mg of PF5%600BODIPY4NH₂, 0.2 mg of PF5TBT4NH₂ and 1.4 mgof PF46BT-4NH₂ were dissolved in 2 mL of THF, then 125 μL (4000 ppm) of0.5 PSMA-8000 (33% MA, 67% PS, Mw: 8000) in THF was added into the aboveTHF solution. The solution was covered with foil to protect it fromlight and stirred for 72 h at room temperature. 100-200 μL of abovesolution was diluted to 40-50 ppm with THF and was used for Pdotspreparation. FIG. 42D shows the absorption and fluorescence spectra ofresulting crosslinked Pdots. The crosslinked 655 Pdots emission exhibita FWHM of 36 nm.

Cross-linking reaction of PF5%680BODIPY4NH₂, PF46BT4NH₂ and PF5TBT4NH₂with PSMA. 0.7 mg of PF5%680BODIPY4NH₂, 0.3 mg of PF5TBT4NH₂ and 1.0 mgof PF46BT-4NH₂ were dissolved in 2 mL of THF, then 125 μL (4000 ppm) of0.5 PSMA-8000 (33% MA, 67% PS, Mw: 8000) in THF was added into the aboveTHF solution. The solution was covered with foil to protect it fromlight and stirred for 72 h at room temperature. 100-200 μL of abovesolution was diluted to 40-50 ppm with THF and was used for Pdotspreparation. FIG. 42A shows the absorption and fluorescence spectra ofresulting crosslinked Pdots. The crosslinked 680 Pdots emission exhibita FWHM of 44 nm.

Example 20 Synthesis of a Narrow-band Emissive Pdots with AbsorptionPeak Near 488 nm

The present example provides a method for obtaining narrow-band emissivepolymer dots with absorption peak near 488 nm (FIGS. 43 and 44A-C).

Synthesis of Compound 2 (FIG. 43). In a 100-mL flask, compound 1 (4 g, 8mmol), KOAc (4 g) and DMF (60 mL) were added. After the mixture wasdegassed twice, Pd(dppf)₂Cl₂ (300 mg) was added. Then, the mixture washeated to 90° C. overnight. The resulted dark solution was poured intowater after cooling to room temperature, and extracted withdichloromethane three times. The organic phase was washed with water anddried over anhydrous Na₂SO₄. After removing the solvent, the crudeproduct was purified by silica column to give a colorless liquid (2.7 g,62%). ¹H NMR (CDCl_(3, δ):) 10.06 (s, 1H), 7.86-7.83 (d, 4H), 7.79-7.76(d, 2H), 2.06 (t, 4H), 1.40 (s, 12H), 1.20-1.15 (m, 24H), 1.01 (t, 6H).Synthesis of compound 3.4-Bromo-7-(bromomethyl)-[2,1,3]-benzothiadiazole (2.2 g, 7.1 mmol) wasadded into a 50-mL flask. After adding triethyl phosphate (15 mL), thesolution was refluxed for 4 hours. Then, the excessed triethyl phosphatewas removed under vacuum to give the crude compound 3, which was usedwithout further purification. ¹H NMR (CDCl₃, δ): 7.84 (d, 1H), 7.53 (m,1H), 4.10 (q, 4H), 3.75 (s, 1H), 3.68 (s, 1H), 1.25-1.20 (t, 7H).

Synthesis of Compound 4 (FIG. 43). Compound 2 (1.2 g, 2.2 mmol) and 3 (1g, 2.7 mmol) in THF (10 mL) were charged into a 50-mL single-neck flaskunder nitrogen flow. The mixture was stirred at room temperatureovernight after t-BuOK in methanol (3 mL) was added dropwisely at 0° C.Then, the solution was poured into water and extracted withdichloromethane twice. The combined organic phase was dried overanhydrous Na₂SO₄. After removing solvent, the crude product was purifiedby silica column to give a yellow solid (0.6 g, 38%). ¹H NMR (CDCl₃, δ):8.09 (d, 1H), 7.90-7.83 (m, 2H), 7.78-7.72 (m, 3H), 7.68-7.61 (m, 4H),2.05 (t, 4H), 1.42 (s, 12H), 1.06 (m, 24H), 0.81 (t, 6H).

Synthesis of PFVBT (FIG. 43). In a 25-mL flask, compound 4 (150 mg, 0.20mmol) was added under nitrogen. Toluene (3 mL), aqueous Na₂CO₃ (2 mL,2M) and A336 (2 drops) were added consequently. The solution wasdegassed twice before adding Pd(PPh₃)₄ (8 mg). Then, the solution washeated to 120° C. for 48 hours. After cooling to room temperature, thesolution was poured into methanol. The resulted solid was dissolved intotoluene and passed through a short column. The concentrated solution wasthen poured into methanol. The solid was collected by filtration anddried under vacuum overnight (90 mg, 82%). ¹H NMR (CDCl₃, δ): 8.23-7.62(m, 10H), 2.14 (br, 4H), 1.15 (br, 24H), 0.82 (t, 6H).

Synthesis of polymer PFVBT-BODIPY (FIG. 43). Here is an example of PFVBTpolymer with 2% BODIPY, named as PFVBT-BODIPY2. In a 25-mL flask,compound 4 (203 mg, 0.27 mmol) and compound 5 (3.4 mg, 0.0055 mol) wereadded under nitrogen. Toluene (4 mL), aqueous Na₂CO₃ (2 mL, 2M) and A336(2 drops) were added consequently. The solution was degassed twicebefore adding Pd(PPh₃)₄ (10 mg). Then, the solution was heated to 120°C. for 48 hours, and phenyl boronic acid (50 mg) was added and stirredfor 12 hours, and bromobenzene (0.2 mL) was added and stirred for 12hours. After cooling to room temperature, the solution was poured intomethanol. The resulted solid was dissolved into toluene and passedthrough a short column. The concentrated solution was then poured intomethanol. The solid was collected by filtration and dried under vacuumovernight (110 mg, 74%). ¹H NMR (CDCl₃, δ): 8.18-7.72 (m, 10H), 2.15(br, 4H), 1.15 (br, 24H), 0.82 (br, 6H).

Preparation and characterization of PFVBT-BODIPY Pdots (FIGS. 44A-C).PFVBT-BODIPY2 Pdots were prepared by nano-coprecipitaiton method. Asolution of polymer blend PFVBT-BODIPY (200 ppm) and PSPEG or PSMA (100ppm)) in THF (4 mL) was quickly injected into water (10 mL) underultrasonication. THF was evaporated by N₂ flow at 95° C. and thesolution was concentrated to ˜8 mL, followed by filtration through a 0.2micron filter. The prepared Pdots aqueous solutions were kept inrefrigerator for further use. The particle size and zeta-potentials ofPdots in bulk solution was characterized by dynamic light scattering(DLS) (Malvern Zetasizer NanoS). UV-Vis absorption spectra were recordedwith a DU 720 scanning spectrophotometer (Beckman Coulter, Inc., CA,USA) using 1 cm quartz cuvettes. Fluorescence spectra were obtainedusing a commercial Perkin-Elmer fluorometer. Fluorescence quantum yieldswere measured using a Hamamatsu photonic multichannel analyzer C10027equipped with CCD integrating sphere.

Example 21 Synthesis of BODIPY Based Narrow-band Emissive Polymer (Shownin FIG. 46) and Blended Narrow-band Emissive Polymer Dots Emitting inthe Near Infrared Region

The present example provides a method for obtaining BODIPY basednarrow-band emissive polymer (FIG. 46) and narrow-band emissive polymerdots emitting in the near infrared region by blending several types ofsemiconducting polymers.

Synthesis of monomer C (FIG. 47A). A mixture of 2,7-dibromofluorene (15mmol, 4.86 g), tert-butyl 3-bromopropanoate (33 mmol, 6.86 g), sodiumhydroxide solution (40%, 35 mL), and Bu₄NBr (1.5 mmol, 0.48 g), toluene(70 mL) was stirred at 85° C. overnight. The organic phase wasseparated, washed with water and dried over MgSO₄. After evaporation ofthe solvent, the residue was purified by column chromatography (DCM).The product was obtained as a white solid. Yield: 4.81 g, 83%. ¹HNMR(500 MHz, CDCl₃): δ=7.47-7.54 (m, 6H), 2.30 (t, 4H), 1.47 (t, 4H), 1.33(s, 18H). ¹³CNMR (500 MHz, CDCl₃): 172.71, 150.47, 139.60, 131.56,126.99, 122.57, 121.93, 80.97, 54.58, 34.92, 30.36, 28.52.

Synthesis of polymer P1 (FIG. 47A). To a 100 mL flask was added monomerA (0.9 mmol, 493 mg), monomer B (1.02 mmol, 569 mg), monomer C (0.1mmol, 58 mg), Bu₄NBr (15 mg), toluene (20 mL), Na₂CO₃ (2M, 10 mL). Themixture was stirred at room temperature and the flask was degassed andrecharged with N₂, which was repeated four times before and afteraddition of Pd(PPh₃)₄ (0.02 mmol, 23 mg). The reactants were stirred at90° C. for 48 hours and then phenylboronic acid (100 mg) dissolved inTHF (1 mL) was added. After two hours, bromobenzene (1 mL) was added andfurther stirred for 3 hours. The mixture was poured into methanol (200mL). The precipitate was filtered, washed with methanol, water, andacetone to remove monomers, small oligomers, and inorganic salts. Thecrude product was dissolved in DCM (15 mL), filtered through a 0.2 μmmembrane and re-precipitated in methanol (150 mL). The powder was thenstirred in acetone (200 mL) overnight and collected by filtration, anddried in vacuum. Yield: 580 mg (74%). ¹HNMR (500 MHz, CDCl₃): δ=7.84 (d,J=7.8 Hz, 2H), 7.63-7.73 (m, 4H), 2.12 (broad, 4H), 1.32 (s, 1H),1.10-1.25 (m, 20H), 0.68-0.92 (m, 10H).

The t-Butyl protected polymer (200 mg) was dissolved in DCM (20 mL) andTFA (3 mL) was added and the mixture was stirred at room temperature indark overnight and then poured into methanol (150 mL). The precipitatewas collected by filtration, washed with methanol, water, and acetonethoroughly, and then dried in vacuum. (¹HNMR and ¹³CNMR).

Synthesis of polymer P2 (FIG. 47B). P2 was synthesized bycopolymerization of monomers 2,7-dibromo-9,9-bis(3-(tert-butylpropanoate))fluorene (A), 9,9-dioctylfluorene-2,7-diboronic acidbis(1,3-propanediol) ester (B), 4,7-dibromobenzo[c][1,2,5]thiadiazole(D) by Suzuki Coupling with different monomer feed ratios. We usePFBT-C2 as an example here: In a 100 mL flask, monomer A (0.18 mmol,104.8 mg), B (0.82 mmol, 235.9 mg), C (1 mmol, 558.4 mg), was dissolvedin toluene (30 mL), Bu₄NBr (0.04 mmol, 12.5 mg) and Na₂CO₃ (2M, 12 mL)was added. The mixture was degassed and refilled with N₂ (repeated 4times) before and after addition of Pd(PPh₃)₄ (0.035 mmol, 40 mg). Thereactants were stirred at 90° C. for 40 hours and phenylboronic acid(100 mg) dissolved in THF (1 mL) was added. After two hours,bromobenzene (1 mL) was added and further stirred for 3 hours. Themixture was poured into methanol (200 mL). The precipitate was filtered,washed with methanol, water, and acetone to remove monomers, smalloligomers, and inorganic salts. The crude product was dissolved in DCM(15 mL), filtered through a 0.2 μm membrane and re-precipitated inmethanol (150 mL). The powder was then stirred in acetone (200 mL)overnight and collected by filtration, and dried in vacuum. Yield: 412mg (70%). ¹HNMR (500 MHz, CDCl₃): δ=7.90-8.20 (m, 8H), 2.00-2.30 (broad,4H), 1.32 (s, 0.86H), 1.08-1.26 (m, 20H), 0.96 (broad, 4H), 0.81 (t, d=6Hz, 6H). The protecting tert-butyl esters group was removed by TFA atroom temperature. Trifluoroacetic acid (3 mL) was added into a solutionof polymer (200 mg) in DCM (60 mL) and stirred overnight. The organiclayer was washed with water (100×3) and then stirred with NaOH solution(10%, 30 mL) for 10 minutes. The mixture was then acidified with aceticacid. The DCM phase was washed with water and concentrated to 10 mL andprecipitated in methanol (100 mL). The final powder was collected byfiltration, washed with acetone, and dried in vacuum.

Synthesis of polymer P3 (FIG. 47C). To a 100 mL flask was added monomerA (0.7 mmol, 384 mg), monomer B (1.02 mmol, 569 mg), monomer E (0.3mmol, 134 mg), Bu₄NBr (15 mg), toluene (20 mL), Na₂CO₃ (2M, 10 mL). Themixture was stirred at room temperature and the flask was degassed andrecharged with N₂, which was repeated four times before and afteraddition of Pd(PPh₃)₄ (0.02 mmol, 23 mg), respectively. The reactantswere stirred at 90° C. for 48 hours and then phenylboronic acid (100 mg)dissolved in THF (1 mL) was added. After two hours, bromobenzene (1 mL)was added and further stirred for 3 hours. The mixture was poured intomethanol (200 mL). The precipitate was filtered, washed with methanol,water, and acetone to remove monomers, small oligomers, and inorganicsalts. The crude product was dissolved in DCM (15 mL), filtered througha 0.2 μm membrane and re-precipitated in methanol (150 mL). The powderwas then stirred in acetone (200 mL) overnight and collected byfiltration, and dried in vacuum. Yield: 532 mg (71%). ¹HNMR (300 MHz,CDCl₃): δ=7.84 (d, J=8.1Hz, 2H), 7.53-7.62 (broad, 4H), 7.48 (t, J=7.2Hz, 0.28H), 7.36-7.43 (broad, 1.07H), 2.12 (broad, 4H), 1.10-1.22 (m,20H), 0.76-0.89 (m, 10H).

Synthesis of polymer P4 (FIG. 47D). To a 100 mL flask was added monomerB (0.122 mmol, 71.7 mg), monomer F (0.022 mmol, 21 mg), monomer G (0.1mmol, 64.2 mg), toluene (4 mL), Na₂CO₃ (2M, 3 mL), ethanol (0.7 mL). Themixture was stirred at room temperature and the flask was degassed andrecharged with N₂, which was repeated four times before and afteraddition of Pd(PPh₃)₄ (0.004 mmol, 4.6 mg), respectively. The reactantswere stirred at 83° C. for 30 hours and then phenylboronic acid (20 mg)dissolved in THF (0.5 mL) was added. After two hours, bromobenzene (0.5mL) was added and further stirred for 2 hours. The mixture was pouredinto methanol (100 mL). The precipitate was filtered, washed withmethanol, water, and acetone to remove monomers, small oligomers, andinorganic salts. The crude product was dissolved in DCM (5 mL), filteredthrough a 0.2 μm membrane and re-precipitated in methanol (60 mL). Thepowder was collected by filtration, and dried in vacuum. Yield: 252 mg(77%). ¹HNMR (300 MHz, CDCl₃): 7.55-7.95 (m, 6.66H), 4.06 (t, J=6.9 Hz,0.31H), 2.09 (broad, 3.65 H), 1.85 (m, 0.32 H), 1.4-0.6 (m, 30H).

Preparation, characterizations and bioapplications of the Pdots (FIG.48-52). The semiconducting polymer dots were prepared byco-precipitation method. All polymers were dissolved into anhydrous THF,respectively, to form a 1 mg/mL THF solution, then as the weight ratioof 50:50:70:8 to mix polymers P1, P2, P3, P4 (for 4-NIR Pdots) THFsolution, and the weight ratio of 100:50:6 to mix polymers P2, P3, P4(for 3-NIR Pdots) THF solution. Then 0.2 mL mixed solution was addedinto 1.8 mL anhydrous THF, which was further injected directly into 10mL DI H₂O under ultrasonication. THF was removed by N₂ flow at roomtemperature. The sizes of these two kinds of Pdots (4-NIR Pdots and3-NIR Pdots) three polymers were characterized as 15.7 nm by dynamiclight scattering. FIG. 48A shows a TEM image of 4-NIR Pdots(P1/P2/P3/P4). FIG. 48B shows particle size distributions of narrow-bandemissive 4-NIR Pdots (P1/P2/P3/P4). FIG. 48C shows TEM image of 3-NIR(P2/P3/P4) Pdots. FIG. 48D shows particle size distributions ofnarrow-band emissive 3-NIR (P2/P3/P4) Pdots. The data were measured bydynamic light scattering. FIG. 49 shows the absorption spectra of 4-NIRPdots (solid line), 3-NIR Pdots (dashed line) and fluorescence spectraof 4-NIR Pdots (dot line, λ_(ex)=380 nm) and 3-NIR Pdots (dashed-dot-dotline, λ_(ex)=450 nm) with narrow emission bands (FWHM=55 nm). FIG. 50Ashows the single-particle fluorescence image of Qdot705. FIG. 50B showsthe single-particle fluorescence image of -NIR Pdots. FIG. 50C shows thecorresponding intensity distribution histograms of Qdot705. FIG. 50Dshows the corresponding intensity distribution histograms of 3-NIRPdots. From the distribute curves of single-particle brightness ofseveral thousand particles of Qdot705 and 3-NIR Pdots, respectively, wecan know that 3-NIR Pdots were 3 times brighter than Qdot 705. FIG. 51Ashows the flow cytometry measurements of the intensity distributions ofMCF-7 cells labeled with 4-NIR Pdot-streptavidin (negative labeling, dotline; positive labeling, solid line). FIG. 51B shows the flow cytometrymeasurements of the intensity distributions of MCF-7 cells labeled with3-NIR Pdot-streptavidin. (negative labeling, dot line; positivelabeling, solid line). All the negative and positive labelings werecompleted and measured under identical experimental conditions, only inthe negative labeling primary biotinylated antibody was absent. FIG. 52shows fluorescence images of MCF-7 breast-cancer cells labeled with3-NIR Pdot-streptavidin. Negative labeling performed under the samecondition but in the absence of the biotinylated primary antibody doesnot show fluorescence signal. From left to right: blue fluorescence fromthe nuclear stain Hoechst 34580; red fluorescence images from 3-NIR-SAprobes; Nomarski (DIC) images; and combined fluorescence images. Scalebars: 20 μm.

Example 22 Narrow-band Emissive Lanthanide Polymer Dots with a <10 nmFWHM Bandwidth Emission

This example describes polymer dots that include a violet-emissivepolymer as a donor and red emissive europium complexes as acceptorsalong with the application of time-gated fluorescence bioimaging basedon the Eu complexes/PVK Pdots. As described further below, the europiumcomplexes were integrated with polymers and condensed to form polymerdots that exhibited a less than about 10 nm FWHM emission.

FIG. 53 shows example polymers and lanthanide complexes that were usedto produce example polymer dots of the present invention. ExampleEuropium complexes included Eu15F and EuDNM. The polymers included,e.g., polyvinyl-N-carbazole (PVK), polystyrene polymer conjugated withpolyethylene glycol having a COOH terminus (PS-PEG-COOH), andpolystyrene (PS). The length of the various polymers can be denominatedby m and n as shown in FIG. 53. The number of repeating units, m and n,can be any desired length. For example, in PVK, n can range from 5 to10,000; in PS-PEG-COOH, m can range from 10-1000, and n can range from1-100; and in PS, n can range from 5 to 10,000). In this example, PVKhas an average Mw=75,000, polydispersity=2. PS-PEG-COOH has a main chainMw=8,500, graft chain Mw=1,200, and total chain Mw=21,700,polydispersity=1.25). PS has an average Mw=41,000.

FIG. 54 shows an example preparation method of making polymer dotscontaining PVK and Eu complex.

Spectroscopic and TEM characterizations of Eu15F/Polymer Pdots wereperformed. FIG. 55A shows that the emission spectrum of PVK overlappedwith the absorption spectrum of Eu15F. This spectral overlap makes itpossible for the fluorescence energy transfer from PVK donor to theacceptor Eu15F. The average particle diameter of the Eu15F (60 wt %)/PVKPdots was about 16 nm as measured by high resolution TEM (FIG. 55B). Thequantum yields of Eu15F (at different ratios)/PVK Pdots were measuredand compared to those of Eu15F in non-fluorescent polymer polystyrene,Eu15F (at different ratios)/PS nanoparticles. As shown in the quantumyield curves (FIG. 55C), self-quenching for these particles occurredwhen the ratio of Eu15F was higher than 60%.

FIG. 56 shows that fluorescence resonance energy transfer (FRET) existedbetween conjugated polymer PVK and Eu15F complex. As shown in theemission spectra (FIG. 57), the intensity of the narrow emission peak ofEu15F complex at 612 nm (characteristic emission of Eu) was increasedwhen the ratio of Eu15F was increased. As shown, the emission intensity(355 nm-500 nm) of PVK dropped when the ratio of Eu15F increased from 0%to 60%. The emission intensity (575 nm-625 nm) of Eu15F increased. Whilenot being limited by any particular theory, the change was likely due tothe energy transfer from PVK to Eu15F. The emission spectra indicatesthat the FWHM of the emission spectra of Eu15F (60 wt %)/PVK Pdots islower than ˜10 nm.

Fluorescence emission intensity from Eu was enhanced at least in part bythe large absorption cross-section of the conjugated polymer PVK, ascompared to PS. FIG. 57 shows emission intensity comparisons betweenEu15F/PS nanoparticles and Eu15F/PVK Pdots with varying Eu15F ratios(e.g., from 20% to 80% Eu15F) upon excitation at 342 nm. As shown inFIG. 57, the emission intensities of Eu15F/PVK Pdots were larger thanthat of Eu15F/PS nanoparticles before the ratio of Eu15F reached 60 wt%. Considering that quantum yields of Eu15F/PS nanoparticles were largerthan that of Eu15F/PVK Pdots when the ratio of Eu15F was in the rangefrom 20%-90% (FIG. 55), the emission intensity enhancement resultsdemonstrated that the absorption cross-sections of Eu15F/PVK Pdots werelarger than that of Eu15F-PS nanoparticles. When the ratio of Eu15Fincreased, self-quenching also began to happen. The self-quenchingeffect became more and more dominant when the ratio of Eu15F wasincreased. For example, when the ratio reached 80%, the emissionbrightness enhancement of Eu15F/PVK Pdots over that of Eu15F/PSnanoparticles decreased to 1.

Spectroscopic and TEM characterizations of EuDNM/Polymer Pdots were alsoperformed. FIG. 58A shows absorption spectra of EuDNM/PS nanoparticles,EuDNM/PVK Pdots and pure PVK Pdots, and the emission spectra of PVKPdots. FIG. 58B shows TEM image of EuDNM/PVK Pdots. FIG. 58C shows thequantum yields of EuDNM/PS nanoparticles and Eu15F/PVK Pdots versus theratios of EuDNM complexes. In this example, the emission spectrum of PVKshows a better overlap with the absorption spectrum of EuDNM than thatof Eu15F. So there can be more efficient energy transfer from PVK toEuDNM complex. The average particle size (diameter) of prepared Pdotswas around 17 nm as measured by high resolution TEM. The quantum yieldsof EuDNM/PVK were measured and compared to that of EuDNM innon-fluorescent polystyrene nanoparticles. As shown in the quantum yieldcurve, decrease of quantum yield due to self-quenching happened when theratio of EuDNM was beyond 50%. The molecular weight of PS was chosen tobe close to that of conjugated polymer PVK.

FIG. 59 depicts emission intensity comparisons between EuDNM/PSnanoparticles and EuDNM/PVK Pdots with varying ratios of EuDNM complex.As shown in FIG. 59, the emission intensities of EuDNM/PVK Pdots werelarger than that of EuDNM/PS nanoparticles when the ratio of EuDNM wasfrom 20% to 80%. In this example, the brightness enhancement ofEuDNM/PVK over EuDNM/PS nanoparticles was much larger than their quantumyield enhancements. The results demonstrated that the absorptioncross-sections of EuDNM/PVK Pdots were larger than that of EuDNM/PSnanoparticles.

This example also includes application of Eu/PVK Pdots for flowcytometry and time-gated fluorescence imaging. The scheme in FIG. 60shows the bioconjugation and cellular surface labeling procedures forEu/PVK Pdots. The bottom curves show the flow cytometry measurements ofthe intensity distributions of MCF-7 cells labeled with Eu/PVK Pdots(negative labeling, left line; positive labeling, right line). All thenegative and positive labeling experiments were completed and measuredunder identical conditions, except that biotinylated antibody was absentin the negative labeling. It is notable, given the long lifetime of theLanthanide Pdots, that they produce excellent signal to noise in theflow cytometry application where signal integration time is so short. Tocollect time-gated fluorescence images, a high speed optical chopper wasplaced in the intermediate image plane of an inverted Nikon T-2000microscope. One focusing lens was placed in the middle of the opticalchopper and the EMCCD camera. The distance between optical chopper andEMCCD was 4 times of the focusing length of the focusing lens. A UV LED(365 nm) was used as the illumination light source. The optical chopperand the UV LED were synchronized through a home-built synchronizer.Time-gated-fluorescence image of Eu15F/PVK Pdots and commercial redfluorescence emitting nanoparticles R300 are shown in FIG. 61. Thefluorescence lifetime of Eu15F/PVK Pdots was measured to be around 500μs; while that of the commercial R300 nanoparticles was around 1 ns. Asshown in images, the Eu15F/PVK Pdots still emitted many photons after200 μs delay (the time between excitation and signal collection). Butthe commercial R300 nanoparticles did not emit photons any more after200 μs delay. The experimental conditions are as below: time delay,around 200 μs; excitation LED wavelength, 365 nm; emission collectionfilter, 545-625 nm; objective, 20×; N.A., 0.65; EMCCD: photonmax;exposure time, 200 ms; image size, 16 μm×16 μm.

The long lifetime of Eu15F/PVK Pdots can be differentiated fromcommercial R300 nanoparticles by utilizing their fluorescence lifetimedifference. As shown in the fluorescence emission spectra (FIG. 62A),both particles emit red fluorescence. Due to the spectral overlap, thetwo types of particles were not differentiated by simply using bandpassfilters (FIG. 62B). However, the two types of particles could be easilydifferentiated by utilizing their lifetime difference. As shown in thenormal fluorescence image (FIG. 62B), both particles emitted photons.When there was time delay between excitation and signal collection, onlysome of the particles emitted photons (FIG. 62C). And these particleswere Eu15F/PVK Pdots. Therefore the two different types of particleswith overlapping emission spectra were easily differentiated by usingtheir difference in lifetime.

Eu15F/PVK Pdots were applied for live cell imaging, as shown in FIG. 63.Eu15F/PVK Pdots were incubated with MCF-7 cells overnight. Before beingplaced on a microscope stage for imaging, the cells were fixed withparaformaldehyde for 10 mins, and then washed with 1×PBS buffer solutionthree times. The experimental conditions were as below: Time delay: 200μs; excitation LED wavelength: 365 nm; emission collection filter: 500nm longpass filter; Objective: 20×; N.A.: 0.65; Exposure time: 200 ms(EMCCD from photonmax); Image size: 85 μm×64 μm. In a normalfluorescence image, the background intensity level is usually high dueto the auto-fluorescence of cells (top left image), while the backgroundintensity level decreased significantly in a time-gated fluorescenceimage (top right image). As a result, the signal/noise ratio could beimproved.

What is claimed is:
 1. A polymer dot comprising: a condensedchromophoric polymer comprising a semiconducting polymer, wherein thesemiconducting polymer comprises a narrow-band emissive monomer, thecondensed chromophoric polymer having an emission spectrum with a fullwidth half maximum (FWHM) of less than about 70 nm, and wherein theratio of the number of narrow-band emissive monomers to the number ofother monomers is about 1:20 or less.
 2. The polymer dot of claim 1,wherein the chromophoric polymer is a homopolymer or a heteropolymer. 3.The polymer dot of claim 1, wherein the chromophoric polymer furthercomprises a first general monomer, a second general monomer, or acombination thereof, and wherein the first and second general monomerhave an emission spectrum with a full width half maximum of greater than70 nm.
 4. The polymer dot of claim 1, wherein the narrow-band emissivemonomer, the first general monomer, the second general monomer, or acombination thereof are integrated into the backbone of the chromophoricpolymer.
 5. The polymer dot of claim 1, wherein the narrow-band emissivemonomer is a BODIPY derivative.
 6. The polymer dot of claim 5, whereinthe BODIPY derivative has the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A) and R^(4B) isindependently selected from the group consisting of hydrogen, alkyl,aralkyl, aryl, and alkoxy-aryl, and wherein the BODIPY derivative isintegrated into the chromophoric polymer by attachment to R¹, R^(2A),R^(2B), R^(3A), R^(3B), R^(4A), R^(4B) or a combination thereof.
 7. Thepolymer dot of claim 1, wherein the narrow-band emissive monomer is asquaraine derivative.
 8. The polymer dot of claim 7, wherein thesquaraine derivative has the formula:

each of X¹ and X² is independently selected from the group consisting ofoxygen, sulfur and nitrogen; each of R^(1A) and R^(1B) is independentlyselected from the group consisting of, but not limited to, alkylene,alkenylene, arylene, heteroarylene, phenylene, azulene, cycloalkylene,and heterocycloalkylene; and each of R^(2A) and R^(2B) is a reactivegroup independently selected from the group consisting of, but notlimited to, a halide, hydroxyl, and amino, and wherein the squarainederivative is integrated into the chromophoric polymer by attachment toR^(1A), R ^(1B), R^(2A), R^(2B) or a combination thereof.
 9. The polymerdot of claim 1, wherein the narrow-band emissive monomer is a metalcomplex derivative.
 10. The polymer dot of claim 1, wherein thenarrow-band emissive monomer is a porphyrin derivative.
 11. The polymerdot of claim 1, wherein the narrow-band emissive monomer is ametalloporphyrin derivative.
 12. The polymer dot of claim 1, wherein thenarrow-band emissive monomer is a lanthanide complex derivative.
 13. Thepolymer dot of claim 1, wherein the narrow-band emissive monomer is aperylene derivative.
 14. The polymer dot of claim 1, wherein thenarrow-band emissive monomer is a cyanine derivative.
 15. The polymerdot of claim 3, wherein the first general monomer and the second generalmonomer are each independently selected from the group consisting offluorene, a fluorene derivative, phenylene vinylene, a phenylenevinylene derivative, phenylene, a phenylene derivative,benzothiadiazole, a benzothiadiazole derivative, thiophene, a thiophenederivative, carbazole fluorene, and a carbazole fluorene derivative. 16.The polymer dot of claim 1, wherein the condensed chromophoric polymerfurther comprises a functional group rendering the surface of thecondensed chromophoric polymer available for conjugation orbioconjugation.
 17. The polymer dot of claim 16, wherein the functionalgroup is a hydrophobic functional group.
 18. The polymer dot of claim17, wherein the hydrophobic functional group is selected from the groupconsisting of alkyne, strained alkyne, azide, diene, alkene,cyclooctyne, and phosphine.
 19. The polymer dot of claim 16, wherein thefunctional group is a hydrophilic functional group.
 20. The polymer dotof claim 19, wherein the hydrophilic functional group is selected fromthe group consisting of carboxylic acid or salts thereof, amino,mercapto, azido, aldehyde, ester, hydroxyl, carbonyl, sulfate,sulfonate, phosphate, cyanate, and succinimidyl ester.
 21. The polymerdot of claim 16, wherein the functional group is conjugated to abiomolecule.
 22. The polymer of claim 21, wherein the biomolecule isselected from the group consisting of a protein, a glycoprotein, apeptide, an amino acid, a metabolite, a drug, a toxin, a nucleic acid, acarbohydrate, a sugar, a lipid, and a fatty acid.
 23. The polymer dot ofclaim 1, further comprising a polymer physically blended or chemicallycross-linked with the chromophoric polymer, wherein the polymer is aconjugated polymer or a non-semiconducting polymer.
 24. The polymer dotof claim 1, wherein the FWHM is less than about 60 nm, less than about50 nm, less than about 40 nm, less than about 30 nm, or less than about20 nm.
 25. The polymer dot of claim 1, wherein the condensedchromophoric polymer comprises a quantum yield greater than about 10%,greater than about 20%, greater than about 30%, greater than about 40%,greater than about 50%, greater than about 70%, or greater than about90%.
 26. The polymer dot of claim 1, wherein the condensed chromophoricpolymer is stable over a period of greater than about 1 month, greaterthan about 3 months, greater than about 6 months, or greater than about1 year.
 27. The polymer dot of claim 1, wherein the polymer dot is freeof secondary structure comprising a beta-phase secondary structure. 28.A polymer dot comprising: a condensed chromophoric polymer comprising asemiconducting polymer, wherein the semiconducting polymer comprises anarrow-band unit attached to the backbone or a sidechain of the polymer,wherein the narrow-band unit comprises an emission spectrum with a fullwidth half maximum (FWHM) of less than about 70 nm, wherein the backbonecomprises a first general monomer, a second general monomer, or acombination thereof, wherein the narrow-band unit comprises anarrow-band emissive monomer, and wherein the ratio of the number ofnarrow-band emissive monomers to the number of other monomers is about1:20 or less.
 29. The polymer dot of claim 28, wherein the narrow-bandunit is covalently attached to the backbone or the sidechain.
 30. Thepolymer dot of claim 28, wherein the narrow-band unit comprises afluorescent nanoparticle embedded in or attached to the polymer dot. 31.The polymer dot of claim 30, wherein the fluorescent nanoparticle is aquantum dot.
 32. The polymer dot of claim 28, wherein the narrow-bandunit comprises a fluorescent molecule.
 33. The polymer dot of claim 28,wherein the fluorescent molecule is a polymer or a dye molecule.
 34. Thepolymer dot of claim 28, wherein the narrow-band emissive monomer is aBODIPY derivative.
 35. The polymer dot of claim 34, wherein the BODIPYderivative has the formula:

wherein each of R¹, R^(2A), R^(2B), R^(3A), R^(3B), R^(4A) and R^(4B) isindependently selected from the group consisting of hydrogen, alkyl,aralkyl, aryl, and alkoxy-aryl, and wherein the BODIPY derivative isintegrated into the chromophoric polymer by attachment to R¹, R^(2A),R^(2B), R^(3A), R^(3B), R^(4A), R^(4B) or a combination thereof.
 36. Thepolymer dot of claim 28, wherein the narrow-band emissive monomer is asquaraine derivative.
 37. The polymer dot of claim 36, wherein thesquaraine derivative has the formula:

each of X¹ and X² is independently selected from the group consisting ofoxygen, sulfur and nitrogen; each of R^(1A) and R^(1B) is independentlyselected from the group consisting of, but not limited to, alkylene,alkenylene, arylene, heteroarylene, phenylene, azulene, cycloalkylene,and heterocycloalkylene; and each of R^(2A) and R^(2B) is a reactivegroup independently selected from the group consisting of, but notlimited to, a halide, hydroxyl, and amino, and wherein the squarainederivative is integrated into the chromophoric polymer by attachment toR^(1A), R^(1B), R^(2A), R^(2B)or a combination thereof.
 38. The polymerdot of claim 28, wherein the narrow-band emissive monomer is a metalcomplex derivative.
 39. The polymer dot of claim 28, wherein thenarrow-band emissive monomer is a porphyrin derivative.
 40. The polymerdot of claim 28, wherein the narrow-band emissive monomer is ametalloporphyrin derivative.
 41. The polymer dot of claim 28, whereinthe narrow-band emissive monomer is a lanthanide complex derivative. 42.The polymer dot of claim 28, wherein the narrow-band emissive monomer isa perylene derivative.
 43. The polymer dot of claim 28, wherein thenarrow-band emissive monomer is a cyanine derivative.
 44. The polymerdot of claim 28, wherein the first general monomer and the secondgeneral monomer are each independently selected from the groupconsisting of fluorene, a fluorene derivative, phenylene vinylene, aphenylene vinylene derivative, phenylene, a phenylene derivative,benzothiadiazole, a benzothiadiazole derivative, thiophene, a thiophenederivative, carbazole fluorene, and a carbazole fluorene derivative. 45.The polymer dot of claim 28, wherein the FWHM is less than about 60 nm,less than about 50 nm, less than about 40 nm, less than about 30 nm, orless than about 20 nm.
 46. The polymer dot of claim 28, wherein thecondensed chromophoric polymer comprises a quantum yield greater thanabout 10%, greater than about 20%, greater than about 30%, greater thanabout 40%, greater than about 50%, greater than about 70%, or greaterthan about 90%.
 47. The polymer dot of claim 28, wherein the condensedchromophoric polymer is stable over a period of greater than about 1month, greater than about 3 months, greater than about 6 months, orgreater than about 1 year.
 48. The polymer dot of claim 28, wherein thepolymer dot is free of secondary structure comprising a beta-phasesecondary structure.
 49. A method of making a polymer dot, the methodcomprising: providing a solvent solution comprising a chromophoricpolymer, the chromophoric polymer being in an elongated coil form andcomprising a semiconducting polymer, wherein the semiconducting polymercomprises a narrow-band emissive monomer, wherein the chromophoricpolymer comprises an emission spectrum with a full width half maximum(FWHM) of greater than about 70 nm, and wherein the ratio of the numberof narrow-band emissive monomers to the number of other monomers isabout 1:20 or less; and mixing the solvent solution comprising thechromophoric polymer with a miscible solvent to form a condensedchromophoric polymer, wherein the condensed chromophoric polymercomprises an emission spectrum with a FWHM less than about 70 nm.
 50. Amethod of making a polymer dot, the method comprising: providing asolvent solution comprising a chromophoric polymer, the chromophoricpolymer being in an elongated coil form and comprising a semiconductingpolymer, wherein the semiconducting polymer comprises a narrow-bandemissive monomer, wherein the chromophoric polymer comprises an emissionspectrum with a full width half maximum (FWHM) of less than about 70 nm,and wherein the ratio of the number of narrow-band emissive monomers tothe number of other monomers is about 1:20 or less; and mixing thesolvent solution comprising the chromophoric polymer with a misciblesolvent to form a condensed chromophoric polymer, wherein the condensedchromophoric polymer comprises an emission spectrum with a FWHM lessthan about 70 nm.
 51. The method of claim 49 or 50, wherein thechromophoric polymer is physically blended, chemically crosslinked, orboth with a polymer comprising a general monomer that acts as an energydonor to the narrow-band emissive monomer, the narrow-band emissivemonomer being an energy acceptor.
 52. The method of claim 49 or 50,wherein the condensed chromophoric polymer comprises a quantum yieldgreater than about 10%, greater than about 20%, greater than about 30%,greater than about 40%, greater than about 50%, greater than about 70%,or greater than about 90%.
 53. The method of claim 49 or 50, wherein themixing step includes nanoprecipitation.
 54. The method of claim 49 or50, wherein the condensed chromophoric polymer is formed by an emulsionmethod.
 55. The method of claim 49 or 50, wherein the emulsion method isa mini-emulsion method.
 56. The method of claim 49 or 50, wherein thesolvent solution comprises tetrahydrofuran.
 57. The method of claim 49or 50, wherein the miscible solvent comprises water.
 58. A method ofmultiplex detection with a polymer dot, the method comprising: detectingthe polymer dot with a detector system comprising a filter configured topass a spectrum of light having a full width half maximum (FWHM) of lessthan about 70 nm, wherein the polymer dot comprises a condensedchromophoric polymer comprising a semiconducting polymer, wherein thesemiconducting polymer comprises a narrow-band emissive monomer, thecondensed chromophoric polymer having an emission spectrum with a fullwidth half maximum (FWHM) of less than about 70 nm, and wherein theratio of the number of narrow-band emissive monomers to the number ofother monomers is about 1:20 or less.
 59. The method of claim 58,wherein the FWHM is less than about 60 nm, less than about 50 nm, lessthan about 40 nm, less than about 30 nm, or less than about 20 nm. 60.The method of claim 58, wherein detecting the polymer dot comprisesusing flow cytometry.
 61. A method of making a polymer dot, the methodcomprising: providing a solvent solution comprising a chromophoricpolymer, the chromophoric polymer being in an elongated coil form andcomprising a semiconducting polymer, wherein the semiconducting polymercomprises a narrow-band unit attached to the backbone or a sidechain ofthe polymer, wherein the narrow-band unit comprises an emission spectrumwith a full width half maximum (FWHM) of greater than about 70 nm,wherein the narrow-band unit comprises a narrow-band emissive monomer,and wherein the ratio of the number of narrow-band emissive monomers tothe number of other monomers is about 1:20 or less; and mixing thesolvent solution comprising the chromophoric polymer with a misciblesolvent to form a condensed chromophoric polymer, wherein the condensedchromophoric polymer comprises an emission spectrum with a FWHM lessthan about 70 nm.
 62. A method of making a polymer dot, the methodcomprising: providing a solvent solution comprising a chromophoricpolymer, the chromophoric polymer being in an elongated coil form andcomprising a semiconducting polymer, wherein the semiconducting polymercomprises a narrow-band unit attached to the backbone or a sidechain ofthe polymer, wherein the narrow-band unit comprises an emission spectrumwith a full width half maximum (FWHM) of less than about 70 nm, whereinthe narrow-band unit comprises a narrow-band emissive monomer, andwherein the ratio of the number of narrow-band emissive monomers to thenumber of other monomers is about 1:20 or less; and mixing the solventsolution comprising the chromophoric polymer with a miscible solvent toform a condensed chromophoric polymer, wherein the condensedchromophoric polymer comprises an emission spectrum with a FWHM lessthan about 70 nm.
 63. The polymer dot of claim 1, wherein thechromophoric polymer is a conjugated polymer.
 64. The polymer dot ofclaim 63, wherein the conjugated polymer is a semiconducting polymer.65. The polymer dot of claim 12 or 41, wherein the lanthanide complexderivative comprises a metal selected from the group consisting ofCe(III), Pr(III), Nd(III), Sm(III), Sm(II) Eu(III), Eu(II), Tb(III),Dy(III), Ho(III), Er(III), Tm(III), Yb(III) and Yb(II).
 66. The polymerdot of claim 65, wherein the lanthanide complex comprises an anionicligand and/or a neutral ligand that chelates to the metal.
 67. Thepolymer dot of claim 1, wherein the narrow-band emissive monomer isselected from a rhodamine, a rhodamine derivative, a coumarin, acoumarin derivative, a xanthene, and a xanthene derivative.
 68. Thepolymer dot of claim 1, further comprising a conjugated polymerphysically blended or chemically cross-linked with the chromophoricpolymer.